PMID- 19463905
OWN - NLM
STAT- MEDLINE
DCOM- 20091110
LR  - 20131121
IS  - 1873-7544 (Electronic)
IS  - 0306-4522 (Linking)
VI  - 162
IP  - 4
DP  - 2009 Sep 15
TI  - The generation of an array of clonal, immortalized cell models from the rat 
      hypothalamus: analysis of melatonin effects on kisspeptin and 
      gonadotropin-inhibitory hormone neurons.
PG  - 1134-40
LID - 10.1016/j.neuroscience.2009.05.026 [doi]
AB  - Significant information on reproductive function has been generated based on the 
      rat model, including many seminal discoveries. Yet little is known about the 
      molecular and cellular events involved in control of reproductive function, 
      mainly due to the pervasive lack of cell models from rat. We have therefore 
      generated a wide array of cell lines using primary cell culture from the rat 
      hypothalamus. Immortalization of the primary cells was achieved through 
      retroviral transfer of T-antigen, followed by selection with geneticin. The mixed 
      cell populations were subcloned and each clonal cell line was analyzed for 
      expression of specific cellular markers. Each line has a distinct phenotypic 
      profile, with expression of key neuroendocrine markers. We have functionally 
      analyzed two clonal cell lines, rHypoE-7 and rHypoE-8, for hormones implicated in 
      the control of gonadotropin-releasing hormone neuronal function through 
      melatonin, specifically kisspeptin (KISS) and RF-amide-related peptide-3 (RFRP-3, 
      the mammalian ortholog of the avian gonadotropin-inhibiting hormone, GnIH). We 
      detected functional melatonin receptor activity, as each cell line exhibited 
      inhibition of forskolin-stimulated 3'-5'-cyclic adenosine monophosphate (cAMP) 
      accumulation. Upon treatment with 10 nM melatonin, we found that KISS gene 
      expression was decreased in the rHypoE-8 cell line, while RFRP-3 was increased in 
      the rHypoE-7 cell line. These results are in accordance with the differential 
      regulatory functions of these two peptides, particularly on GnRH neuronal 
      control. These cell lines will serve as novel tools for the analysis of the 
      cellular and molecular mechanisms involved in hypothalamic control of a number of 
      physiological processes described in the rat animal model.
FAU - Gingerich, S
AU  - Gingerich S
AD  - Department of Physiology, University of Toronto, Medical Sciences Building 3247A, 
      1 King's College Circle, 3247A MSB, Toronto, ON, Canada M5S 1A8.
FAU - Wang, X
AU  - Wang X
FAU - Lee, P K P
AU  - Lee PK
FAU - Dhillon, S S
AU  - Dhillon SS
FAU - Chalmers, J A
AU  - Chalmers JA
FAU - Koletar, M M
AU  - Koletar MM
FAU - Belsham, D D
AU  - Belsham DD
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090520
PL  - United States
TA  - Neuroscience
JT  - Neuroscience
JID - 7605074
RN  - 0 (Kiss1 protein, rat)
RN  - 0 (Kisspeptins)
RN  - 0 (Neuropeptides)
RN  - 0 (Proteins)
RN  - 0 (RFamide peptide)
RN  - 0 (RNA, Messenger)
RN  - 0 (Receptors, Melatonin)
RN  - 33515-09-2 (Gonadotropin-Releasing Hormone)
RN  - JL5DK93RCL (Melatonin)
SB  - IM
MH  - Animals
MH  - *Cell Line
MH  - *Clone Cells
MH  - Embryo, Mammalian/cytology
MH  - Gonadotropin-Releasing Hormone/*antagonists & inhibitors
MH  - Hypothalamus/*cytology
MH  - Kisspeptins
MH  - Melatonin/pharmacology/*physiology
MH  - Neurons/cytology/*metabolism
MH  - Neuropeptides/genetics/*metabolism
MH  - Proteins/*metabolism
MH  - RNA, Messenger/metabolism
MH  - Rats
MH  - Receptors, Melatonin/metabolism
EDAT- 2009/05/26 09:00
MHDA- 2009/11/11 06:00
CRDT- 2009/05/26 09:00
PHST- 2009/04/07 00:00 [received]
PHST- 2009/04/30 00:00 [revised]
PHST- 2009/05/14 00:00 [accepted]
PHST- 2009/05/26 09:00 [entrez]
PHST- 2009/05/26 09:00 [pubmed]
PHST- 2009/11/11 06:00 [medline]
AID - S0306-4522(09)00842-2 [pii]
AID - 10.1016/j.neuroscience.2009.05.026 [doi]
PST - ppublish
SO  - Neuroscience. 2009 Sep 15;162(4):1134-40. doi: 
      10.1016/j.neuroscience.2009.05.026. Epub 2009 May 20.