Immunocytochemical identification and localization of APUD cells in the gut of seven stomachless teleost fishes

Table 1 Details of antisera and main reagents used

Antisera against	Working dilution	Code	Specificity	Source
Synthetic human gastrin	1:5000	GP-1304	No cross reaction with cholecystokinin-8	Dr. N Yanaihara,Shizuoka
Leucine-enkephalin	1:80000	1671	—	UCB-Bioproducts Bruxelles
Bovine pancreatic polypeptide	1:12000	615-R-110	Cross-react s with human pancreatic polypeptide	Dr. RE Chance, Indianapolis
Substance P	1:2000	MAS 035B	—	Sera-Lab., Sussex
Gastric inhibitory polypeptide	1:10000	G/R/34-IIID	No cros s reaction with glucagon	Dr. D Grube, Hannove
Porcine glucagon	1:1000	RPN1602	Wholly cross react with pancreatic	Amersham International pl.,
			and intestinal glucagon	Amersham
Synthetic human cyclic
somatostatin	1:3000	—	—	Dr. S Ito Niigata
Glucagon-like immunoreactants	1:1000	RPN1604	Wholly cross react with pancreatic	Amersham International pl.,
			and intestinal glucagon	Amersham
Insulin	1:1000	47291	—	—
Avian pancreatic polypeptide	1:10000	Iance-10/5/81	No cross reaction with glucagon	Dr. JR Kimmel, Kansas City
5-Hydroxytriptamine	1:10000	Lot.16302	—	Immunonuclear Corp.Stillwater
Synthetic porcine motilin	1:1000	R-1104	Reacts against entire molecules	Dr. N Yanaihara, Shizuoka
Natural porcine
cholecystokinin-33	1:3000	—	Reacts with cholecystokinin 11-20,	Dr. J Yamada, Shizuoka
			no cross reaction with gastrin
Synthetic porcine secretin	1:1000	R-801	Reacts with the C- and N-terminals	Dr. N Yanaihara, Shizuoka
Synthetic bovine neurotensin	1:1000	R-3501		Dr. N Yanaigar a, Shizuoka
Synthetic porcine vasoac tive
intestinal polypeptide	1:2000	R-502	Reacts against entire molecules	Dr. N Yanaihara, Shizuoka
Bombesin	1:3000	27070	Cross reaction with GRPImmunonrclear	Corp. Stillwater
Neuron specific enolase	1:1000	—	—	Dako, Copenharge
Prochymosin	1:2000	—	—	A. Andren, Uppsala
Pepsinogen	1:2000	—	—	A. Andren, Uppsala
Rabbit PAP	1:100	Z - 113	—	Dako, Copenharge
Guineapig PAP	1:50	24699	—	Dako, Copenharge
PAP Kit	1:100	61- 2003	—	Zymed Lab. Inc., SouthSan
				Francisco, USA
ABC Kit	1:50	PK-4001	—	Vector Lab. Burlingame USA

Table 2 Distribution and density of 8 IR endocrine cells in the gut of three teleost species (^-x ± s)

Fish species	Gut parts	GAS	ENK	B P P	SP	GLI	GIP	GLU	SOM
Crass carp	I	64 ± 13	85 ± 17	52 ± 12	56 ± 14	44 ± 19	23 ± 19	61 ± 7	—
(C.idellus)	II	26 ± 6b	57 ± 11a	31 ± 16c	76 ± 20a	45 ± 22c	11 ± 7	44 ± 4b	—
	III	—	22 ± 7b	—	57 ± 10c	27 ± 9c	—	31 ± 3b	—
	IV	—	17 ± 10b	—	17 ± 9b	—	—	21 ± 5b	—
	V	—	1 ± 2b	—	18 ± 7b	—	—	9 ± 5b	—
Black carp	I	77 ± 8	57 ± 6	72 ± 6	20 ± 5	68 ± 17	59 ± 13	2 ± 3	—
(M. piceus)	II	63 ± 5b	51 ± 5c	58 ± 7a	47 ± 4b	88 ± 11c	34 ± 20a	1 ± 5c	—
	III	32 ± 4b	38 ± 4b	22 ± 4b	38 ± 3b	92 ± 30c	31 ± 19a	—	—
	IV	23 ± 5b	57 ± 9c	—	60 ± 7b	16 ± 12b	17 ± 11b	—	—
	V	—	86 ± 12b	—	72 ± 9b	10 ± 6b	8 ± 6b	—	—
Common carp	I	25 ± 6	38 ± 4	12 ± 6	—	—	4 ± 3	2 ± 2	13 ± 12
(C. carpio)	II	20 ± 5c	39 ± 5c	9 ± 8c	18 ± 4b	6 ± 1b	2 ± 2c	1 ± 5c	8 ± 5 b
	III	12 ± 2b	28 ± 3b	—	31 ± 6b	11 ± 4b	—	—	10 ± 8 b
	IV	3 ± 4b	9 ± 5b	—	19 ± 3b	—	—	—	—
	V	—	—	—	—	—	—	—	—

I = Anterior segment of foregut; II = Posterior segment of foregut; III = M idgut; IV = Anterior segment of hindgut; V = Rectum; —: IR endocrine cell wasn’t found, PAP method was used in the table; n = 5,

^aP < 0.05,

^bP < 0.01,

^cP > 0.05.

Table 3 Distribution and density of 3 IR endocrine cells in the gu t of four teleost species (^-x ± s)

Fish species	Gut parts	GAS	GIP	GLU
Silver carp	I	17 ± 5	2 ± 2	43 ± 10
(H. molitrix)	II	2 ± 4b	6 ± 3a	64 ± 35c
	III	—	—	126 ± 36b
	IV	—	—	61 ± 26c
	V	—	—	30 ± 28c
Bighead	I	35 ± 10	13 ± 3	29 ± 16
(A. nobilis)	II	—	—	27 ± 7c
	III	—	—	79 ± 7b
	IV	—	—	60 ± 62c
	V	—	—	47 ± 46c
Silver crucian	I	32 ± 17	1 ± 2	53 ± 18
Carp (C. gibelio)	II	3 ± 3b	—	20 ± 11b
	III	—	—
	IV	—	—	11 ± 13b
	V	—	—	—
Bluntnose black	I	31 ± 8	4 ± 1	32 ± 16
Bream (M.	II	—	—	24 ± 6c
Amblyocephala)	III	—	—	4 ± 3b
	IV	—	—	24 ± 11c
	V	—	—	6 ± 8b

I = Anterior segment of foregut; II = Posterior segment of foregut; III = Midgut; IV=Anterior segment of hindgut; V = Rectum; —: IR endocrine cell wasn’t found, ABC method was used in the table; n = 5,

^aP < 0.05,

^bP < 0.01,

^cP > 0.05.