The effects of [D-Ala6,Pro9-NEt]-LHRH (LHRH-A) alone and in combination with drugs which influence the actions of dopamine or the synthesis of catecholamines on gonadotropin (GtH) secretion and ovulation in the loach (Paramisgurnus dabryanus) were investigated. LHRH-A alone stimulated an increase in serum GtH levels in the loach, but was a relatively ineffective treatment for the induction of ovulation. Injection of the dopamine receptor antagonist pimozide caused a marked potentiation of the GtH-release response to LHRH-A, and combined injections of pimozide and LHRH-A were an effective treatment for the induction of ovulation. Reserpine, a drug which causes depletion of catecholamines from presynaptic terminals, also caused a marked potentiation of the GtH-release response to LHRH-A and combined treatment induced ovulation. Similarly, administration of alpha-methyl-para-tyrosine to block conversion of tyrosine to L-dopa, or carbidopa to block conversion of L-dopa to dopamine, potentiated the GtH-release response to LHRH-A and induced ovulation. In ontrast, the use of diethyldithiocarbamate, to block conversion of dopamine to norepinephrine, failed to augment the action of LHRH-A on GtH release and ovulation. The present results provide further vidence to suggest that dopamine functions as a gonadotropin release-inhibitory factor in teleosts, and demonstrate that the use of drugs which block either the synthesis or the actions of dopamine potentiates the action of LHRH-A in teleosts.

(1) In tadpoles, chicken-II gonadotropin-releasing hormone (cGnRH-II) could be measured in the brains before metamorphosis, but mammalian gonadotropin-releasing hormone (mGnRH) did not appear until the stage of metamorphosis, i.e. cGnRH-II appeared earlier than mGnRH during ontogenesis. (2) During the metamorphic climax, mGnRH content increased more rapidly than cGnRH-II; the content of mGnRH was about two times of that of cGnRH-II. (3) In juveniles and adults, the content of mGnRH and cGnRH-II, and the distribution pattern of mGnRH (but not cGnRH-II) in the brains and pituitaries changed with age and stages of gonadal development. mGnRH mainly distributed in the rostral brain areas, whereas cGnRH-II had a widespread distribution in the brain. (4) Both mGnRH and cGnRH-II were present in the pituitaries at each stage of maturity. The gonadotropin-releasing hormone (GnRH) content at sexually maturity was significantly higher than that at other stages of gonadal development, and the content of mGnRH was about 15-18 times of that of cGnRH-II. (5) These results suggest that both mGnRH and cGnRH-II are potentially involved in the direct regulation of pituitary gonadotropes, and mGnRH may be the major active form, cGnRH-II may also serve as a neurotransmitter or neuromodulator in the brain.

Effects of cysteamine hydrochloride (CSH)-a somatostatin-inhibiting agent on growth hormone (GH) secretion from pituitary fragments (PF) or hypothalamus plus pituitary fragments (HPF) under static incubation conditions, serum GH, 3,5,30-triiodothyronine (T3) and thyroxine (T4) levels, and growth in juvenile grass carp (Ctenopharyngodon idellus) were investigated. CSH (0.1, 1, and 10mM) had no influences on GH release from PF after 1 and 6h incubation, but was effective in stimulating GH release from HPF in a dose-dependent manner after 1 and 6h incubation. Moreover, prolonged treatment of HPF with CSH decreased the magnitude of enhancement of GH levels in culture medium. CSH and neuropeptides [e.g., human GH-releasing hormone (hGHRH, 100nM), luteinizing hormone-releasing hormone analog (LHRH-A, [D-Trp6, Pro9]LHRH, 100 nM)], or salmon gonadotropin-releasing hormone analogue (sGnRH-A, [D-Ala6, Pro9]LHRH, 100 nM), alone and in combination during static incubation stimulated GH release from HPF after 1h incubation; in addition, there was an additive, not a synergistic effect of CSH and neuropeptides on stimulation of GH release. Administration of CSH (2.5mg/g diet) in combination with LHRH-A (5lg/g diet) in diet twice daily for 8 weeks resulted in higher serum GH, T3, and T4 levels, ratio of RNA/DNA in muscle, food conversion efficiency, and growth rate than CSH or LHRH-A alone. At trial termination, significant decreases in condition factors and body lipid levels were observed in fish fed with CSH and/or LHRH-A. No significant differences were recorded for viscero-somatic index, hepato-somatic index, and percent body moisture and protein in muscle. These findings, taken as a whole, strongly suggest that the action of CSH stimulating GH release in vitro appears to be mediated through hypothalamic pathways and dietary delivery of CSH directly or indirectly stimulates endogenous GH, T3, and T4 secretion, and subsequently leads to a increase in growth rate in grass carp.

A full-length cDNA encoding the neuropeptide Y (NPY) was cloned from the hypothalamus of orange spotted grouper (Epinepheluscoioides) by rapid amplification of cDNA ends approaches. The NPY cDNA sequence is 688 bp long and has an open reading frame of 300 bp encoding prepro-NPY with 99 amino acids. The deduced amino acid sequences contain a 28-amino-acids signal peptide followed by a 36- amino-acids mature NPY peptide. mRNA expression of NPY was determined using semi-quantitative RT-PCR followed by Southern blot analysis. NPY mRNA was expressed in olfactory bulb, telencephalon, pituitary, hypothalamus, optic tectum-thalamus, medulla oblongata, cerebellum and spinal cord. Low levels of NPY mRNA expression were found in retina, ovary and stomach, while much lower levels of expression were detected in liver, heart, gill, skin, anterior intestine, thymus and blood. No NPY mRNA expression was observed in unfertilized eggs, newly fertilized eggs, 16-cells stage and morula stage of the embryo and lower levels of expression were detected in the blastula, gastrula and neurula stages. It was highly expressed from lens formation stage to 52-day-old larval stage. NPY might be involved in the late embryonic and larval development of the orange spotted grouper.