File Id: ABS/PB/MOL/6 CODE:Molecular

Analysis of daily and circadian gene expression in the rat pineal gland
Fukuhara C. and Tosini G. (Morehouse School of Medicine, USA)
Neuroscience Research Volume 60, Issue 2, February 2008, Pages 192-198

The mammalian pineal gland is an important component of the circadian system. In the
present study, we examined the expression of roughly 8000 genes in the rat pineal gland
as a function of time of day under light–dark (LD) cycles and in constant dark (DD) using
oligo DNA microarray technique. We identified 47 and 13 genes that showed higher
levels at night and day, respectively, under LD. The same patterns of expression were
also observed in DD. About half of the genes that peaked at night have a known
biological function, i.e., transcription factors and proteins that are involved in signaling
cascades, whereas 14 are expressed sequence tags and 8 have an unknown biological
function. Twelve of the genes that were up-regulated at night were also up-regulated after
1 h NE stimulation, thus suggesting that the expression of these genes is controlled by
adrenergic mechanisms. Of the 13 genes that were up-regulated in the daytime, 6 coded
for proteins that are involved in intracellular signaling pathways. The results obtained
with microarray analysis were well correlated with data obtained using real time
quantitative RT-PCR. The present results provide new materials to dissect and understand
the pineal physiology.

File Id: ABS/PB/light CODE:Tosini

Effect of Long-Term Exposure to Constant Dim Light on the Circadian System of

Rats
Fukuhara C., Aguzzi J., Bullock N. and Tosini G.
Neurosignals 2005;14:117–125

The newly discovered multi-oscillatory nature of the mammalian circadian clock system
and the cloning of the genes involved in the molecular mechanism that generates
circadian rhythmicity have opened new approaches for understanding how mammals are
temporally organized and how the mammalian circadian system reacts to the lack of
normal synchronization cues. In the present study we investigated the effects of long-
term exposure to constant red dim light on the pattern of the expression of Period 1 in the
suprachiasmatic nuclei of the hypothalamus and of Arylalkylamine N-
acetyltransferase(Aa-nat) in the retina and pineal gland. Our data demonstrate that
Period 1 mRNA expression in the suprachiasmatic nuclei of the hypothalamus was not
affected by exposure to constant red dim light for 60 days, whereas Aa-nat mRNA
expression in the retina and in the pineal gland was significantly affected, since in some
animals (20-30%) Aa-nat mRNA levels were found to be higher during the subjective
day. A circadian rhythm of serum melatonin and locomotor activity was present in all the
animals tested. In 4 animals serum melatonin levels were high during the subjective day.
Our data suggest that long-term exposure to constant red dim light may induce
desynchronization between the circadian rhythm of locomotor activity and serum
melatonin levels.
File Id: ABS/PB/MOL/7 CODE:Molecular/PER

Regulation of Period 1 Expression in Cultured Rat Pineal

Chiaki Fukuhara, James C. Dirden, Gianluca Tosini
Neurosignals, Vol. 11, No. 2, 2002

The aim of the present study was to investigate the in vitro expression of Period 1 (Per1),
Period 2 (Per2) and arylalkylamine N-acetyltransferase (AA-NAT) genes in the rat pineal
gland to understand the mechanism(s) regulating the expression of these genes in this
organ. Pineals, when maintained in vitro for 5 days, did not show circadian rhythmicity in
the expression of any of the three genes monitored. Norepinephrine (NE) induced AA-
NAT and Per1, whereas its effect on Per2 was negligible. Contrary to what was observed
in other systems, NE stimulation did not induce circadian expression of Per1. The effect
of NE on Per1 level was dose- and receptor subtype-dependent, and both cAMP and
cGMP induced Per1. Per1 was not induced by repeated NE - or forskolin - stimulation.
Protein synthesis was not necessary for NE-induced Per1, but it was for reduction of Per1
following NE stimulation. Per1 transcription in pinealocytes was activated by
BMAL1/CLOCK. Our results indicate that important differences are present in the
regulation of these genes in the mammalian pineal.

File Id: ABS/PB/Mus terricolor

Chromosomal and molecular divergence in the Indian pygmy field mice Mus
booduga-terricolor lineage of the subgenus Mus
Sharma T.
Genetica 97: 331-338, 1996

Mus booduga and Mus terricolor both have 2n=40. Unlike M. booduga, with all
acrocentric chromosomes, M.terricolor invariably has large submetacentric X and
acrocentric Y due to an increase of heterochromatin. In contrast to the conservative
karyotype of the co-existing sibling species booduga, three chromosome types of
terricolor are found in different populations and their divergent karyotypes have
autosomal heterochromatin variations established in the homozygous condition. The
average genetic distance determined from electrophoretic study of 20 protein loci ranges
from lowest (D=0.106) between chromosome types I & II to highest (D=0.185) between
types II & III. In terricolor, booduga and M. m. tytleri high mean values of variations per
locus (range A---- 1.604 to 1.928) and heterozygosity per individual per locus (range
H=0.180 to 0.336) have been observed. Sequence divergence of 0.39 to 1.2%, calculated
from restriction profiles of mtDNA, shows that the terricolor chromosome types have
diverged recently. Hybridizations between type I females and type III males gave a
preponderance of males in the F1 with varying degrees of sterility. The 'terricolor
complex' is an interesting system for critical probing for the role of heterochromatin in
the process of speciation. MtDNA, protein loci and AT-rich musculus-related major and
minor satellite DNA data indicate that progenitors of the booduga-terricolor lineage
might have evolved simultaneously with the caroli-cookii-cervicolor lineage in the
evolution of the subgenus Mus.