A tutorial in basic neurobiology

byDavid L. Atkins
Professor of Biology
George Washington University
This file loads many drawings...be patient.
Contents of this step:
o eview Pores
o !mportant !ons
o "ydrated !ons
o !oni# elative $i%es
o !on $i%e&Pore $i%e
o !oni# 'on#entrations
o (le#tri#al Gradients
o $odi)m&Potassi)m P)mp
o *ernst (+)ation and...
o ...Donnan (+)ation
o Goldman (+)ation
Review Pores
By way of review, it is ass)med that yo)
)nderstand basi# #ell membrane str)#t)re
. emember that protein pores e-tend thro)gh the plasma membrane. These
vary in si%e and in n)mber so that a variety of mole#)lar&ion si%es #an be transported
passively a#ross the membrane in n)mbers depending on pore distrib)tion and
#on#entration&ele#tri#al gradients.
!f yo) are following the proper se+)en#e of #hapters, we have dis#)ssed the #ell
membrane str)#t)re, barriers to diff)sion a#ross those membranes, and ways aro)nd
those barriers. We have talked abo)t osmosis..diff)sion a#ross a semi.permiable
membrane.
!n Step 2 we shall dis#)ss ele#tri#al barriers, ele#trolytes, ele#troni# pores /gates0,
and the development of ele#tri#al gradients a#ross the membrane.
Important ions
Animal #ells e-ist in a /relatively0 #ontrolled environment of ions..even the great ma1ority of
marine organisms whi#h are isotoni# to their environment, live in the largest ioni# b)ffer in
the known )niverse..the o#ean2. 3therwise, some #ombination of )rinary system and ion
p)mps at gill, digestive, skin, et#. s)rfa#es maintains a stable internal milea). The ma1ority of
work on ne)rons has been done on three animal gro)ps, and we4ll )se these as o)r working
e-amples..s+)id, frog, and mammals /mainly seven spe#ies, abo)t half being primates0.
Anyway, the ions #on#erned are potassi)m /5
6
0, #hloride /'l
.
0, sodi)m /*a
6
0, and #al#i)m
/'a
66
0. These are listed in order of in#reasing si%e of the hydrated ion, whi#h be#omes
important when yo) learn that most pores are large eno)gh to pass 5
6
, b)t few will #arry *a
6
,
and almost none #an transport 'a
66
.
2 $in#e all living #hemistry and physi#s o##)r in water,
yo) sho)ld ass)me that all ions are in their hydrated
state.
Hyrate
ions
Water mole#)les are asymmetri#al, the hydrogen atoms lying at 789: to ea#h other. This
prod)#es a slightly polari%ed mole#)le that is attra#ted to other polari%ed mole#)les or to ions.
This bond is weak, and in effe#t an ion is simply handed off thro)gh a series of water
mole#)les. The water #lo)d is part of the whole. ;or an ion to move thro)gh the lipid bilayer,
it wo)ld have to shed its asso#iated waters, a pro#ess so e-pensive energeti#ally that it simply
does not happen. The waters of hydration vs. the hydrophobi# lipid layers prevents movement
of ele#trolytes a#ross the #ell membrane. Pores an gates are necessary!
Please note that the n)mber of water mole#)les shown in these drawings is meant to be
symboli# depi#tion rather than a##)rate #o)nt. The a#t)al n)mber of water mole#)les
for any ion spe#ies depends on the energeti# state of the sol)tion at that time.
Ionic relative si"es
The #lo)d of water mole#)les that asso#iates with an ion is somewhat indeterminant in si%e,
depending on energy dynami#s of the sol)tion. This is irrelevant in a living system sin#e all
hydrated ions are otherwise in the same sol)tion..their absol)te and relative si%es are in an
environment so stable that the hydrated ioni# diameters will always be #onsistent <not
4#onstant4= and predi#table. 'ells #an adapt to those si%es with repeatable s)##ess. "owever,
the larger the )nhydrated ion in si%e, the more dispersed is its own #harge and the less
strongly it will attra#t water. This leads to the in#ongr)o)s sit)ation of the larger the ion, the
less hyrate it is, and the smaller the relative si"e of the hyrate whole. Th)s, 5
6
> *a
6
,
b)t hydrated 5
6
? hydrated *a
6
.
Ion si"e#pore si"e
The fig)re at left shows the sit)ation generally fo)nd in #ell membranes /e-#ept the pores are
m)#h more s#attered0. There is a relative ab)ndan#e of pores large eno)gh to pass 5
6
2 b)t too
small for *a
6
. There is a small pop)lation of
#hloride pores, and a small n)mber of *a
6
and 'a
66
pores. The effe#t of this, of #o)rse, is a differential
permiability for these @ ions a#ross the membrane.
A)ite simply, there are very many more entryways
for 5
6
than for sodi)m ion, and very many more *a
6

passages than #al#i)m ion t)nnels. The intra#ell)lar
ion pop)lations refle#t these differen#es.
Ionic concentrations
The #hart at right shows the event)al distrib)tion of the #ommon ions a#ross the plasma
membrane..the steady state.2 'al#i)m ion is not reported be#a)se its e-tra#ell)lar
#on#entration is very low, and its intra#ell)lar presen#e is almost %ero. "owever, the non.
diff)sible anion, A
.
, is in#l)dedB it has a profo)nd infl)en#e on the transmembrane ion
distrib)tion. The A
.
is symboli# of a h)ge s)ite of #ompo)nds whi#h, be#a)se of their
mole#)lar si%e, insol)bility, or bo)nd position in the #ytoplasm, #annot migrate a#ross the
membrane. Cost of these are proteins, polypeptides, organophosphates /s)#h as D*A, *A,
n)#leotides s)#h as ATP, and an array of n)trients. They all behave as organi# a#ids, giving
off a hydrogen ion /that is in#orporated with o-ygen to form metaboli# water0 and leaving the
negative ion inside the #ell.
This establishes a negative #harge dispersed thro)gho)t the
#ytoplasm, and positive ions are attra#ted. 3nly 5
6
pores are
#ommon, so potassi)m ion #on#entrates within the
#ell...sodi)m ion remains o)tside. Apply a bit of simple math
to the #hart to see that total #on#entration inside&o)tside is in
balan#e, and that the #ell is ele#tri#ally ne)tral.

2 $teady state DE e+)ilibri)mF (+)ilibri)m e-ists when
sol)tions on both sides of the membrane are e-a#tly balan#ed
and remain soB steady state indi#ates that both #ompartments
are appro-imately balan#ed, b)t there is a #ontin)ing state of
fl)-.


$lectrical graients
3n the pre#eeding s#reen we #onsidered the effe#t of the #on#entration gradient on
distrib)tion of ob1e#ts /ions0 a#ross a semipermiable membrane. !f that system is allowed to
progress to steady state, the #on#entration of ob1e#ts on both sides of the membrane
/osmolarity0 will balan#e. The distrib)tion of ioni# #harge on these ob1e#ts #ompli#ates the
relationship.
The osmolarity #hart above shows this #ompli#ation. (a#h of the ions has a )nit #harge
e-#ept A
.
. e#all that this is a#t)ally a #ompli#ated array of organi# and impermiant inorgani#
anions. Cost bear a single negative #harge, b)t some are .G and a small n)mber are .@. The
#omposite #harge for the entire array is A
.7.G.

This e-plains a +)estion whi#h sho)ld have o##)rred to yo)..even tho)gh the plasmalemma
is less permiable to sodi)m ion, some sodi)m pores do e-ist, and given eno)gh time the
#on#entrations of sodi)m and potassi)m ions, inside and o)tside, sho)ld be e+)al. ;a#toring
in ioni# #harges yields a very different dynami#.
;irst, sin#e potassi)m #an penetrate the membrane so m)#h easier than sodi)m, the
intra#ell)lar distrib)tion of potassi)m and sodi)m ions sho)ld appear +)i#kly /b)t then
grad)ally alter toward e+)ality0. B)t now we see
positive potassi)m ion being attra#ted to a
negative interior, and th)s reinfor#ing its greater
permiability. Potassi)m ion movement balan#es
the osmolarity e+)ation and does so along an
ele#tri#al gradient of negative inside. As the
potassi)m ioni# #on#entration in#reases, an
in#reasing #on#entration gradient opposes the
ele#tri#al gradient )ntil the two are balan#ed
/steady state0.
A similar intera#tion e-ists between the negative
#ytoplasm and the #on#entration of tiss)e fl)id
#hloride. The hydrated #hloride ion is m)#h
smaller that the hydrated sodi)m ion, and there
are many more membrane pores that will
a##omodate it. (le#tro.rep)lsion of the A
.
works
against the #on#entration gradient of 'l
.
to the
same e-tent as attra#tion for the 5
6
.
The balan#ed osmolarity on both sides of the membrane to some e-tent relieves the
individ)al #on#entration gradients of the several ions. "owever, none of these passive fa#tors
wo)ld maintain the *a
6
&5
6
ratio alone. There is also a dynami# for#e.

Soium#potassium Pump
3ne protein #omponent of #ell membranes is an en%yme, sodi)m.potassi)m.dependent
ATPase. /70 An a#tive site of the en%yme on the inside of the membrane a##)m)lates *a
6
. /G0
When @ sodi)m ions are atta#hed, a se#ond a#tive site /the ATPase0 splits an ATP ..> ADP 6
P
i
6 e. /@0 The energy released reshapes the en%yme mole#)le to move the @ sodi)ms to the
o)tside of the membrane. /H0 This new physi#al form of the mole#)le no longer will IholdI
sodi)m ion, and the e ions are now released o)tside the #ell. This #hange #reates an affinity of
the mole#)le for 5
6
. /J0 When G of these have been a##)m)lated, the mole#)le snaps into a
new shape, moving the potassi)m ions to the inside of the #ell membrane. Again, this final
form is instable, and the potassi)m is released, re#reating the original p)mp mole#)le /70.
Additional intra#ell)lar sodi)m, e-tra#ell)lar potassi)m and ATP will repeat the #y#le.
Any )ne+)al e-#hange of 5
6
for *a
6
is #orre#ted by movement of 5
6
inward thro)gh
membrane pores along its ele#tri#al gradient.

Summary% so far
Th)sK
7. !oni# si%es varyK /A
.
0 >> /*a
6
0 > /'l
.
0 > /5
6
0.
G. Pore si%es and fre+)en#y varyK
/5
6
0 pores, very many
/'l
.
0 pores, many
/*a
6
0 pores, few
/A
.
0 pores, none
Therefore, 5
6
permiability is high /arbitrarily defined asL70, 'l
.
permiability is intermediate
/8.M, relative to potassi)m standard permiability0, *a
6
permiability is low /8.7, relative to 5
6
0,
and A
.
permiability is %ero.
@. (a#h of the above ions has a )nit #harge e-#ept A
.
/.7.G0. A##)m)lation and isolation of A
.

inside the #ell prod)#esK
a. A strong ele#tri#al gradient o)tside.to.inside for 5
6
and *a
6
. "igh penetran#e of 5
6

yields a high inner #on#entration of 5
6
to a steady state of #on#entration gradient vs.
ele#tri#al gradient. Low penetran#e of *a
6
prod)#es low inner #on#entration of *a
6

despite high ele#tri#al and #on#entration gradients.
b. A strong ele#tri#al gradient inside.to.o)tside for 'l
.
. "igh penetran#e of 'l
.
yields a
low #on#entration of 'l
.
inside along its ele#tri#al gradient b)t against its #on#entration
gradient.
H. A metaboli#ally.driven *a&5 p)mp #onstantly removes what sodi)m does enter the #ell,
repla#ing it with e-tra#ell)lar potassi)m. This maintains the dis.e+)ality of 5
6
&*a
6

#on#entrations at a #ontin)ing #ost of ATP.
It is important&&&
...that yo) )nderstand at this point..the A
.
in the #ell is more negative in its #harge than the
#ombined #on#entrations of intra#ell)lar 5
6
and *a
6
. The very high o)tward #on#entration
gradient of 5
6
prevents the entry of more 5
6
. The low penetran#e of *a
6
pl)s its a#tive
removal by the *a
6
&5
6
p)mp blo#ks the development of a high #on#entration of *a
6
inside the
#ell.
'hat this means&&&
...the inside of the living #ell maintains a negative #harge relative to the o)tside.
By convention% the outsie charge of the cell membrane is "ero&
(ernst $)uation
By the end of the 7Nth #ent)ry, it was known that
the #ytoplasm was high in 5
6
and that <*a
6
= was
very low..and that this relationship was reversed
o)tside the #ell. The ass)mption was made that
the #ell membrane was permiable to 5
6
b)t not to
*a
6
. Dire#t meas)rement of the transmembrane
potential was not yet possible, b)t an effort was
made to #al#)late this voltage )sing the *ernst
e+)ation, shown at left.
*onnan $)uilibrium

!mmediately it was )nderstood that #ell membranes were permiable to #hloride as well as to
potassi)m ion, and that these two ions formed an e+)ilibri)m. $in#e both are monovalent ions
b)t of opposite #harge, the Donnan.Gibbs
e+)ilibri)m seemed a good model of the
ele#tri#al transmembrane relationship. The
ass)mptions were permiability and e+)al
#on#entrationsK
................ To prove this to yo)rself, ba#k
)p to the *ernst e+)ation and go solve
identi#al spe#ies and temperat)res for these
two ionsB yo) sho)ld get identi#al answersF
Algebrai# simplifi#ation yieldsK #ard field id G !nversion of the #hloride ion relationship to
remove the min)s sign, pl)s #an#ellation of log on either side prod)#es the Donnan
e+)ilibri)mK #ard field id @ When G ions #an #ross the membrane, the prod)#t of their
e-tra#ell)lar #on#entrations e+)als the prod)#t of their intra#ell)lar #on#entrations.

+olman $)uation
Goldman e+)ation was derived to solve for transmembrane potential )sing all ions involved
sim)ltaneo)sly. ;or most animal #ells, the only important ions are 5
6
, *a
6
, and 'l
.
. $o, the
statement of the e+)ation below lists these @, b)t there is no inherent limitation in the n)mber
of paired #on#entrations. *oti#e also that Goldman in#l)des the permiability fa#tor, p, of ea#h
ion. $ol)tion of the Goldman e+)ation for a #ell membrane yields an a##)rate model of the
transmembrane voltage at any parti#)lar set of #on#entrations and temperat)res.
*oteK sin#e #hloride ion has a #harge opposite to the two #ations, a #orre#tion is needed
to prevent the #ations and anion from #an#elling ea#h other. Th)s, the statement of
relative #hloride ion #on#entrations is inverted..inside over o)tside.
Another noteK sin#e the #on#entrations of potassi)m ion inside L #hloride ion o)tside,
and potassi)m ion o)tside L #hloride ion inside, #orre#ting for the negative #harge on
the #hloride ion...the *ernst of these two ions yields the same answer. Therefore, the
in#l)sion of #hloride ion #on#entrations in Goldman e+)ations is red)ndant. 3mit the
#horide ion, and yo) will get the same answer. Later, when yo) think abo)t a#tion
potentials, yo) will reali%e that only the sodi)m and potassi)m ions migrate a#ross the
membrane in signifi#ant n)mbers, so only they are pra#ti#ally important in
)nderstanding membrane..ion interrelationships.
'hat it all means&&&
Well, reality is not +)ite as straighforward as all that, )nfort)nately...b)t it is #lose. Oo)
sho)ld #arry o)t several #al#)lations of *ernst and Goldman e+)ations to appro-imate the
transmembrane resting potential of #ells. !f so, yo) have seen that the two e+)ations give a
very #lose answer. The reason the two methods are not identi#al is that there really is more
than one ioni# a#tor on this stage, and ea#h is modifying the effe#t of the other.
The reason res)lts of *ernst and Goldman are not f)rther apart is that altho)gh Goldman
in#l)des the modifying fa#tor of ion permiability fa#tors as well as #harge and #on#entration,
the differen#e between the permiabilities of potassi)m and sodi)m is so great we almost have
the *ernst ass)mption..that the membrane is freely permiable to potassi)m and impermiable
to sodi)m. Almost, b)t not +)ite.
There is another #ompli#ation. !t is no longer ne#essary to try to model the resting #ell with
mathemati#s. Codern instr)mentation and te#hni+)es permit dire#t meas)rement. !nsertion of
ele#trodes into the #ytoplasm and a#t)al re#ording of the transmembrane voltage show that the
Goldman #al#)lations are a very good estimate of the tr)th...b)t not e-a#t. Thinking abo)t the
dynami#s of pores and ions makes an )nderstanding of this ine-a#tit)de fairly #lear. $odi)m
penetran#e is P(O small, b)t nonetheless it does e-ist. This means there is a #onstant
leakage of sodi)m inward. $in#e that makes the #ytoplasm slightly less negative, potassi)m
ions have less of an ele#tri#al gradient holding them inside, and they follow their e-#essive
#on#entration gradient o)tward thro)gh membrane pores. The de#rease in internal negativity
also means less rep)lsion of #hloride ions /whi#h we have been ignoring0, and they follow
their #on#entration gradient /and red)#ed ele#tri#al #o)nter.gradient0 into the #ell. The #ell is
now slightly more negative, whi#h attra#ts more sodi)m...b)t...whi#h is also being p)mped
o)tward in e-#hange for slightly fewer potassi)m...

&&&an so on an on&
!n addition, no two cells are e,actly ali-e. There will be variation in the n)mber and si%e of
membrane pores and variation in the e-a#t #omponent and #harge of the A
.
anions lo#ked
within the #ell. This latter will vary #ontin)o)sly with the metaboli# state and a#tivity of the
#ell at any given moment. ;rom these modifiers yo) sho)ld now )nderstand why we speak of
a Isteady stateI rather than an Ie+)ilibri)mI and why the #omple-ities of real #ell life will
skew the real membrane potential slightly from the predi#ted voltage. Goldman is a##)rate...if
the #ell sit)ation wo)ld stay stati#. !t won4t.
There is another point yo) need to )nderstand, altho)gh its #l)es to this point have been too
s)btle to #at#h. !f yo) think ba#k, yo) will re#all that all des#riptions have pointed to Ithe
#ellIB 4ne)rons4 or 4m)s#le #ells4 been mentioned spe#ifi#allyF Transmembrane voltage is a
feat)re of all living animal #ells. !n 'hapter @, we will move on to A'T!3* P3T(*T!AL$ ,
the spe#ial provin#e of nerve and m)s#le fibers. B)t before we do that, finish this review and
be s)re yo) really )nderstand the physi#al and biologi#al basis for the differing distrib)tions
of ions and #harges in the intra#ell)lar and e-tra#ell)lar #ompartments. Give this information
some time to digest. !n#identally, there is some new material in the n)mbered s)mmary
paragraphs belowF

.o review in etail&&&
...any of the points below, #li#k the )nderlined hot link at the bottom of the s#reen. 'li#k the
Iret)rnI b)tton, )pper right of that s#reen, to ret)rn dire#tly to here.
7. The #ell membrane is mainly a bilayer of phospholipid mole#)les, the polar head
e-tending into the e-tra#ell)lar and intra#ell)lar sol)tion. This membrane is b)ttressed
with steroid mole#)les, and s#attered thro)gho)t are many protein mole#)les. $ome of
these are t)b)lar and e-tend a#ross the f)ll thi#kness of the membrane. Water and polar
sol)tes smaller than pore diameter #an e-#hange between the intra. and e-tra#lell)lar
#ompartments.
G. The prin#ipal ions involved in transmembrane potential are potassium% soium%
chlorie% an calcium. *ot m)#h as been said abo)t this latter. ;irst, it is not very
sol)bleB therefore, it is not very ab)ndant. $e#ond, #al#i)m.si%ed pores are e-tremely
rare. Third, #al#i)m dynami#s are different from the other ele#trolytes. !n most #ells
there is no p)mp me#hanism to remove #al#i)m from the #ell, b)t there is a p)mp
whi#h transports #al#i)m ion a#ross the mito#hondrial membraneB inside the
mito#hondrion #al#i)m is )sed in energy metabolism. We shall also dis#)ss two very
spe#ial sit)ations with #al#i)m #on#entrations and movements in the synapse and in
m)s#le #ell f)n#tion. Another point to note is that there is a #hloride p)mp /)s)ally
driven by ATP0 whi#h may be oriented either inward or o)tward and may or may not
be involved with an ion e-#hange. This is a #ompli#ation we #an ignore for the present.
@. Water of hydration of these prin#ipal ions has two effe#ts. !t prevents passage of the
ions thro)gh the hydrophobi# layers of the lipid bilayer. !t magnifies differen#es in ion
diameter so that pore reg)lation of ion movement a#ross the membrane #an be very
pre#ise.
H. These variations in hydrated ion si%e determine whi#h spe#ies of ions will do what 1ob.
J. Potassi)m ion is small eno)gh to pass easily thro)gh a sodi)m pore. *ot vi#e versa. $o,
any biologi#al system whi#h attempted to )se a membrane freely permiable to sodi)m
wo)ld swamp the #ytoplasm with ion to the point of isotoni# distrib)tions of both
sodi)m and potassi)m and #hloride, and the transmembrane potential wo)ld be %ero.
$hortly, yo) will )nderstand that this wo)ld make nerve and m)s#le #ell f)n#tion
impossible. Core to the point, water balan#e wo)ld also be )pset, and in fa#t no #ell
#o)ld s)rvive.
M. Don4t forget the non.diff)sible anion in all of this. !ts non.diff)sion reg)lates the
#ell)lar #on#entration of water and of sol)te ions. Q3smoti# gradients tableR
S. The anion4s #harge determines that those sol)te ions with be #ations and establishes the
ele#tri#al gradient that intera#ts with the #on#entration gradients of those diff)sible
anions and #ations. Q(le#tri#al gradients tableR
9. The system is inherently leaky, and this seepage of *a
6
&5
6
is #orre#ted by the
metaboli#ally driven ion.dependent p)mp . emember that e #onditions are re+)ired
for this p)mp to operate.../a0 there m)st be sodi)m ion in the #ytoplasm, /b0 there m)st
be ATP available to drive the protein mole#)lar re#onfig)ration, and /#0 there m)st be
potassi)m ion for e-#hange in the e-tra#ell)lar environment. Q*a
6
&5
6
p)mp #y#le of
drawingsR
N. The )ltimate res)lt of these ioni# distrib)tions is that ALL animal #ells a##)m)late a
strong negative #harge in their #ytoplasm that is partly offset by an infl)- of potassi)m
ion. elative sodi)m and #hloride ion #on#entrations o)tside the membrane ins)re that
the whole system is ele#tri#ally ne)tral. QDistrib)tion of #harges at #ell membrane
fig)reR
78. The *ernst e+)ation estimates the effe#t on transmembrane potential, given that /a0 the
membrane is permiable to water and to an ion, /b0 the membrane is impermiable to any
other ions in the system, and /#0 time is effe#tively infinite so that e+)ilibri)m will be
established between the two sides of the membrane..th)s, permiability /rate of passage0
of the ion is not a fa#tor.
77. The Donnan .Gibbs e+)ilibri)m establishes that potassi)m on one side of the
membrane is in #on#entration e+)ilibri)m with #hloride /the ele#tri#ally e-#l)ded
anion0 on the other side. $in#e ea#h of these ions has an e+)al b)t opposite #harge, their
polari%ation on either side of the membrane #reates the same transmembrane potential
as well..the e+)ilibri)m is ele#tri#al as well as vol)metri#. ;inally...
7G. The Goldman e+)ation adds the #on#entration effe#ts of all of the ions /a kind of
m)ltipli#ation of the *ernst0 and #orre#ts for the relative permiabilities of ea#h of the
ions /taking potassi)m as the standard, 7.80 so that a real.time&real ele#trolyte estimator
of transmembrane voltage is available. This is not a perfe#t model, b)t it is so #lose that
the minor variations of reality will #an#el to a point of insignifi#an#e.
Well, if yo) really do )nderstand all of this, yo) are ready to begin to think abo)t ne)ron and
m)s#le #ell f)n#tion. ;irst yo) might #he#k yo)rself with the ATA e-er#ise below. .hin-
abo)t any differen#es between yo)r answer and mine.