446

TiPS- December 1991 /Vol. 121

Effccrs (Saxena. I. R. and Fozard, J. R..
eds), pp. 42149, Birkh3user
25 Goadsby, P. J and Edvinsson, L. (1991)
CcphrluI$iIl 11 (Suppl. 11). 3-l
26 Buzzi. M. C. and Moskuwitz. M. A.
(1990) Br. j. fhanr~acol. 99, 202-206
27 Bucklev, T. L.. BrainS. D.. Ramport. M.
and Williams. T I. (1991) Br. 1. Pknr~tie-
col. 103, 1515-1519
28 Anthony, M., Hinterberger, H. and
Lance, J. W. (1%9) R6. C/in. Sfsd.
Headache 2. 29-59
29 Kimball, R. W.. Friedman, A. P. and
V&Jo, E. (1960) Nwrof~y 10,107-111
30 Ogden, H. D. fl%3) Htadathr 3,29-31
31 Lance, J. W. (1973) Mecheabm end
Mnngmwnf of Headechr (2nd edn),
p. 121, Buttenvorths
32 Ostfeld, A. M. and Wolff, H. C. (1955)
Arch. Newel. Ps~yrhiaf. 74, 131-136
33 Schoeffter, P. and Huyer, D. (19B9)
Nmvyt-Srhmitd. Arch. Ph~nttacol. 340,
135-w
34 Humphrey, P. P. A. cf al. fl9BB) Br. J.
Pharmacol. 94.11234132
35 Pemn, M. J., Feniuk, W. and
Humphrey, P. P. A. (1991) Br. 1. Phwma-
rol. 102,191-197
36 Sumner, M. and Humpixey, P. P. A.
(1989) Br. J. Pharmocal. 98.29-31
37 Watts, A. D., Feniuk, W. and
Humphrey, P P. A. (1981) J. Phorm.
PhnrmPcol. 33, SlS-520
Co-secretion of multiple signal molecules
from endocrine cells
via distinct exocytotic pathways
Neurons secrete a cocktail of
neurotransmitter molecules via
both exocytotic and non-exo-
cytotic mechanismslJ. Exocytotic
mechanisms involve at least two
pathways of secretion that use
either synaptic vesicles or large
dense-core vesicles as secretory
organelles (Fig. 1). Synaptic ves-
icles are a characteristic feature of
nerve endings and were con-
sidered until recently to be
neuron-specific organelles. They
contain only non-peptide neuro-
transmitters and play a dominant,
although not exclusive, role in the
fast, point-to-point communi-
cation typical of the nervous sys-
tem. Large dense-core vesicles
contain peptide neurotransmitters
and play an important role in
modulatory signalling. Large
dense-core vesicles may be seen
as the neuronal equivalent of se-
cretory granules of endocrine cells,
which secrete peptide hormones.
Similarities behveen neurons and
peptide-secreting endocrine cells
The biogenesis and biochemical
properties of large dense-core
vesicles and secretory granules are
the same. Many of the same regu-
latory peptides that are secreted
by neurons via large dense-core
vesicles as neurotransmitters are
also secreted by endocrine cells
via secretory granules as peptide
hormones. In addition, a variety
of proteins thought to play a
general role in the organization of
secretory granules (the so-called
granins) are also present in large
dense-core vesicles. Amine
neurotransmitters are present in
secretory granules of certain
endocrine cells and, correspond-
ingly, amines are present in the
large dense-core vesicles of certain
neuron2. In fact, the property to
uptake and decarboxylate amine
precursors has long been recog-
nized as a special characteristic of
neurons and of many endocrine
cells, and led to the concept of the
APUD (amine-precursor uptake
and decarboxylation) neuroendo-
crine system6. The hypothesis that
all cells in this system share the
common embryological origin of
the neural crest has now been
disproven. Yet the special bio-
chemical and functional similarity
of a variety of peptide-secreting
endocrine cells and of neurons has
been strengthened in recent years
and justifies the definition of
this class of endocrine cells as
neuroendocrine cells. Apart from
the contents and the membrane
proteins of large dense-core ves-
icles and secretory granulurs, both
neurons and neuroendocrine cells
express a number of ptokins that
are not expressed by other cells.
Furthermore, a variety of neuro-
endocrine cells have been shown
to express neuronal character-
istics, including that of extending
neurite-like
ptocesses when
grown under certain experimental
conditions. The best-character-
ized example of this phenotypic
shift is the nerve growth factor-
induced neuronal differentiation
of chromaffin cells9. Similar
phenotypic shifts have been dern-
on&rated to occur spontaneously,
or after a variety of experimental
manipulations, in cells and cell
lines derived from other endocrine
tissues~*.
syluptic4ikeu&roveside8
Recent findings suggest the
existence of an additional general
similarity between neurons and
neuroendocrine cells. Following
the identification and character-
ization of several of the major
proteins of synaptic vesicle mem-
branes, it has been found that
many of these proteins (e.g. syn-
aptophysin, ~2% synaptotagmin,
synaptobrevin, SV2, rab3A) are
also expressed at significant
concenhations by subsets of
neuroendocrine cells. Within
TiPS- December 1992 [Vof. 121
neuroendocrine cells they are con-
centrated (and at least some of
them, e.g. synaptophysin and ~29,
are selectively localized) in the
membrane of a population of
microvesicles (synaptic-like micro-
vesides) distinct from secretory
gran~les~*~*~ (Figs 1 and 2). Like
authentic synaptic vesides, syn-
aptic-like microvesides undergo
exocytosis, endocytosis and re-
cyding17-9. One attractive possi-
bility is that the function of
synaptic-like microvesides might
be to store and secrete neuro-
transmitters or similar molecules.
Recent studies carried out on g
cells of the pancreas have sup-
ported this hypothesis. For
many yea? it has been known
that the n euro&msmitter GABA,
as weU as the enzymes involved in
its synthesis (glutamic acid de-
carboxylase) and metabolism
(GABA transaminase), are ex-
pressed at high levels in g cells
of the islets of Langerhans (the
neuroendocrine ceUs that secrete
insuUn)a0. Current evidence
suggests that GABA is secreted
from g cells (which constitute the
core of the i&s) and acts as a
paracrine inhibitory signal mol-
ecule on oerl&e!ral islet cells
(pAmadly ?? g&agon-secreting a
ceW? This view is consistent
wi& the direction of Mood flow in
islets (blood capillaries perfuse
first the g-cell core and then the
peripheral cells of the islets) and
with the antagonistic efkrts of
insulin endplllucagon on glucose
metabolism . Since GABA had
not been deteded in insulin-
containing secretory granules, its
mechanism of secretion remained
undear.
The discovery of synaptic-like
microvesides raised the possi-
bility that GABA secretion from fi
cells might be medfated by these
orgmelles. If synaptic-like micro-
vesides of g cells are involved in
the storage of GABA, they should
share with synaptic vesides of
GABAergic neurons those proper-
ties that are related to GABA up
take and storage. Results obtained
so far using J3 cells in situ and/or
J3-cell lines have shown that this is
the case. First, the GABA syn-
thesizing enzyme glutamic acid
decarboxylase, which in the ter-
minals of GABAergic neurons is
concentrated around synaptic
vesides, was found to be concen-
trated around synaptic-like micro-
vesides of g ceils. Secondly,
regions of g cells enriched in
synaptic-like microvesicles were
also found to be enriched in
GABA immunoreactivity by im-
munofluorescence. Thirdly, a
GABA transport system driven by
a vacua&r proton pump, similer to
the system present in synaptic
vesicles, was present in synaptic-
like microvesicles of J3 cells
(A. Reetx, J. Hell, R. Jahn and
P. De CamiUi, unpublished).
There is no indication that
GABA might be present in syn-
aptic-like microvesicles of neuro-
endocrine cells other than pan-
creatic Jl cells. However, results
obtained with microvesides of J3
cells support the possibility that
synaptic-like microvesicles of
neuroendocrine cells in general
may contain other mdecules
identical or similar to those
that are stored in synaptic ves-
ides in neurons. Neurotrans-
mitter phenotype varies from one
neuron to another. Therefore, a
similar heterogeneity of the neur(l-
transmitter phenotype of endo-
crine cells would not be surpris-
ing. It will be of interest to
determine whether synaptic-like
microvesicles of chromafffn cells
and chromaffin cell-derived cell
lines contain acetylcholine, since
acetylcholine secretion from these
cells has been documentedas. Pre-
liminary evidence obtained in un-
differentiated PC12 cells suggests
that this is the case (T. Flatmark,
A. Regnier-Vigouroux and W.
Huttner, pers. commun.). So far
there is no positive evidence that
synaptic-like microvesicles of
PC12 cells contain catecholiimines.
Growing evidence suggests a
widespread distribution of recep-
tors for non-peptide neurotrans-
mitters outside the nervous sys-
tem. Nemotransmitters secreted
from neuroendocrine cells may
play a role as physiological effec-
tors at such non-neuronel sites.
Synaplicvesidesarenot
mfic
Irrespective of whether synap-
tic-like microvesides of all neuro-
endocrine cells will turn out to
store neurotransmitter-like mol-
ecules and have a signalling func-
tion, synaptic vesicles can no
longer be considered a bona-fide
neuron-specific organelle. They
appear to represent a neuron-
specific adaptation of synaptic-
like microvesicles. A transition
from synaptic-like microvesicles
to authentic synaptic vesides is
recapitulated when neuroendo-
crine cells undergo neuronaf dif-
ferentiation. For example, neur-
onal differentiation of PC12 cei l s
i s accompanied by an increased
expression of the synapsins2, a
class of proteins selectMy associ-
ated with authentic synaptic vet+
ides in nerve terminals3. Clearly,
an important tine of research is
448
the elucidaticn of the mOkCUlaf
mechanisms that CO&~! the
switch from an endocrine to a
neuronal phenotype. Given their
many similarities to synaptic ves-
icles, synaptic-like microvesicles
are a useful model system to study
aspects of the cell biology of syn-
aptic vesicles that cannot be easily
studied in nervous tissue or in
prima8y neuronal cultures. Several
endocrine cell lines are available
in which experiments of genetic
manipulation can be easily per-
formed to study the role of specific
proteins in the exocytotidendo-
cytotic cycling of synaptic vesicles
and synaptic-like microvesicles.
Some new insights into the cell
biology of synaptic vesicles have
already been obtained by the
study of synaptic-like micro-
vesj&+5*7-19.
The finding that synaptic ves-
icles are closely related to an
organelle that is present in neuro-
endocrine c el l s raises the question
of whether the function of both
synaptic vesicles and synaptic-
like microvesicles is in turn some-
how related to the functions of
organ&s present in all cells.
Over the past several years it has
become clear that even the most
specialized cell characteristics rep-
resent adaptations of features ex-
pressed by all cells and can be
seen as the result of a process of
cellular evolution. It is intriguing
that many non-peptide neuro-
transmitters are metabolites in a
variety of metabolic pathways. It
is also intriguing that proteins
that transport metabolites across
membranes (glucose transporters)
are enriched in vesicular carriers,
which, like synaptic vesicles,
undergo exocytotic/endocytotic
recycline. It is possible that early
in cellular evolution metabolic
pathways and signalling path-
ways were closely interrelated.
When the primary sequence of
synaptic vesicle neurotransmitter
carrier proteins is elucidated, it
will be of interest to search for
homologous proteins in micro-
vesicles of non-neuronal, non-
endocrine ceils.
0 Cl cl
Many new and interesting
avenues of research in cellular
endocrinology are opening ahead.
Do synaptic-like microvesicles of
all neuroendocrine cells indeed
contain neurotransmitter-like mol-
ecules? Is exocytosis of synaptic-
like microvesicles regulated? If
so, is exocytosis of synaptic-like
microvesicles and of secretory
granules differentially regulated?
These studies may open new di-
rections in endocrine physiology
and pharmacology. In addition,
they may reveal previously un-
known relationships between
neuronal and endocrine diseases.
PI ETRO DE CAMI LLI
Drpartmrnt of Cell Biology, Yale Uniucrsity
School of Medicine, 333 Cedar Street, New
Huuor. CT 06S?O. USA.
References
1 Hiikfelt, T., Johmsson. 0. and
Gold&in, M. (19B4) Scienct 225,
13264334
2 Garthwaite, J. (1991) Trrnds Neumsci. 14.
60-67
3 De Camilli, P. and Jahn, R. (1990) Annu.
Rrv. Physiol. 52,62?i-645
4 Huttner, W. B., Gerdes, H. H. and Rosa,
P. (1991) Trtnds Biochrm. so. 16, W-W
5 Klein, R., Lagexranh, H. and
Zimmcrmann, H. (1%2) Neumtrms-
miftcr Vesicles, Academk Pmss
6 Pwse, A. C. E. (1977) Med. Biol. 55,
115-125
7 Le Douarin, N. N. (199O) Cell 53,
169-171
Etrata
5-HT advances 011 hr fronts,
by AI&n Abbott (October 1991,
pp. 359-360)
The correct Ki value for
DALI6285 at S-HT, receptors is
185nrbi, not 3nM as erroneously
published.
Ret toMforneump@idey:
mrrl $ pie subtypes and mul-
tiplc second
7 6Y
Martin C. Michcl (Octa et 1991,
pp. 389-394)
Figure 1 should not have been
cited on p. 390. Instead, the
final sentence of the left-hand
column on p. 391 should read
as below.
compound, i.e. C-terminal
fragments and analogues of
NPY (see Fig. l).
We apologize for these errors.
TiPS - December 1991 /Vol. 121
8 Cratzl, M. and La&q K.. eds (1991)
Markers for Ncvnl and Endocrine Ceffs,
VCH
9 Patterson. P. H. (1990) Cefl62.1Q35-1O3B
I O ~e~t2, A.. T. et it. (i990) EMBO 1. 10,
127S-12B4
11 Alema. S., Casalbore, P.. ABostini, E.
and Tato, F. (l!XlS) Nuture 316.557-559
12 Tooze, J., Hoilinahead, M., FuIler, S. D.,
Tome, S. A. and ?iulhw, W. B. (1989)
Eur. 1. Cell Bid. 499.259473
13 Navone, F. et al. (19B6) I. Cell Biot. lO3,
2511-2527
14 Wiedmmann, B., Rehm, H., Knierim,
M. and Becker. C. M. 11988) FEBS Lctt.
240,71-n
15 Cutler, D. F. and Cramer. L. P. (19%)
1. Cell Biol. 110,721~73O
16 Baumert, M. ct PI. (1990) 1. Ccl1 Biol. 110,
12s1294
17 Johnston, P. A. ct al. (19B9) EMBO j. B,
2e4%zB72
18 Cameron, P. L, Sudhof, T.,]ahn, R and
De Cmilli, P. (1991) 1. Cell Btol. 115,
151-164
19 Clift-OGrady, L., Linstedt, A. D.,
Lowe, A. W., Gmte, E. md KeUy, R. 1).
(1990) 1. Cell Bid. llO,l693-17O3
2O Gany, 0. J., Sixensn, R L., Eide, R. P.,
Maley, B. E. end Madsen, A. (19&i)
Di&tes 35,191-l%
21 Ronman, P. et PI. (1989) N&we 341,
22 Banner-Weir, S. and Omi, L. (1983)
Dfrbetes 31, BB3-BB9
23 Schubert, D. and Klkr, F. C. (1977) Prec.
Netf Acad. sci. USA 74.51Bt5lBB
24 Romeno, C., Nichob, R A., Greequd,
P. and Greene, L. A. (1%7) /. Neumscf. 7,
1294-1299
25 .%t, J. W., Ceuze, H. J., C&en@, S.,
Lienhard, C. E. and J ams, D. E. (1991)
1. Cell Biol. 113, 123-236
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