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Mostofa Jamala, Kiyoshi Amenoa, Takanori Mikib, Naoko Tanakaa, Mitsuru Kumihashia and Hiroshi Kinoshitaa 

aDepartment of Forensic Medicine, Faculty of Medicine, Kagawa University, 1750-1, Ikenobe, Miki, Kita, Kagawa 761-0793, Japan 
bDepartment of Anatomy and Neurobiology, Faculty of Medicine, Kagawa University, Japan 


Total text pages are 17 including 2 figures and 1 legend

Address for correspondence
Mostofa Jamal, MD., PhD
Assistant Professor
Department of Forensic Medicine 			
Faculty of Medicine, Kagawa University 
1750-1, Ikenobe, Miki, Kita
Kagawa 761-0793, Japan
Tel: +81-87-891-2140
Fax: +81-87-891-2141
E-mail:  HYPERLINK "mailto:jamaldr68@yahoo.com" jamaldr68@yahoo.com


Abstract
ApoE-knockout (ApoE-KO) mice as a model of apolipoproteinE (ApoE)-deficiency in humans, have been suggested to be involved in the alteration of learning and memory. Our recent study demonstrates a poorer spatial memory in ApoE-KO mice. To further this work, we compared ApoE-KO and C57BL/6J (WT) mice, for expression of choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) mRNA in the hippocampus.  Hippocampal slices were collected and mRNA levels for cholinergic markers were analyzed by real-time PCR. The results revealed a significant strain difference, with ApoE-KO mice demonstrating lower ChAT in the hippocampus compared to WT mice. There were no strain effects on AChE mRNA. Our data provide evidence supporting the role of ApoE in the expression of ChAT and suggest a possible link between cholinergic deficit and poorer performance in ApoE-KO mice. 

Keywords: Hippocampus; Cholinergic markers; ApoE-knockout mice 

Introduction
Choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) are specific cholinergic marker proteins for the functional state of cholinergic neurons, both of which can play a key role in the maintenance of acetylcholine (ACh) levels at the cholinergic neurons [1]. The former catalyzes the biosynthesis of ACh in the cytoplasm of presynaptic terminals, whereas the latter is responsible for inactivating ACh at cholinergic synapses. The cholinergic neurons are involved in several neuropsychic functions such as learning, memory, sleep etc. ACh may play a vast role in modulating these functions [2]. 

Apolipoprotein E (ApoE) is one of the major transporters of cholesterol in the body and is essential for maintaining various neural functions in the brain. For example, ApoE has been suggested to be involved in alterations of brain morphology [3] and cognitive functioning, including attention [4]. Researchers have been shown an impairment of memory associated with cholinergic deficit in ApoE-KO mice [5]. Perturbed cholinergic transmission is involved in number of disease states, including Alzheimer�s disease, Parkinson�s disease, schizophrenia, and autism [6,7,8]. 

Recently, we have shown that ApoE-KO mice have poorer spatial memory [9]. To explore the underlying mechanism, we used ApoE-KO mice, a model of ApoE-deficiency and C57BL/6J (WT) mice to investigate the strain effects on cholinergic markers expression in the hippocampus. To test this, hippocampal slices were collected and the tissue samples for ChAT and AChE mRNA were analyzed by real-time PCR.

2. Materials and Methods
Animals 
All animal experiments were approved by the Kagawa University Animal Investigation Committee. All mice used were of the C57BL/6 genetic background. Breeding pairs for ApoE-KO mice were used to generate the experimental groups. Two 8-week-old male/female pairs of ApoE-KO mice were purchased from the Jackson Laboratory (Bar Harbor, ME) and then bred at the Kagawa University animal facility. The male WT mice used as the controls were purchased from CLEA Japan (Tokyo, Japan). All experiments were conducted with mice that were 10-12 weeks of age and weighed 24-28 g. All animals were housed under controlled temperature (21 � 3 �C), humidity (50-70%), and light (12-h light-dark cycle) conditions. 

Slice preparation
Mice from each strain (ApoE-KO and WT, n = 8 per group) were anesthetized with diethyl ether and then decapitated. Their brains were quickly removed and washed in ice-cold (4�C) artificial cerebrospinal fluid (aCSF) that was saturated with carbogen gas (95% O2, 5% CO2). The aCSF consisted of 124 mM NaCl, 21 mM NaHCO3, 3 mM KCl, 1.25 mM NaH2PO4, 1.6 mM CaCl2, 1.8 mM MgSO4, and 10 mM glucose at pH 7.4. Coronal brain slices (500��m thick) through the hippocampus were cut with a microslicer (DTK-1000, Dosaka, Japan) while immersed in the ice-cold aCSF. The slice was then dip immediately to an oxygenated aCSF solution. Using a standard microscope for visualization, the hippocampus was removed and immediately transferred to a tube containing 0.5 ml of Trizol (Invitrogen). The tissue samples were then homogenized with Polytron (PT 2500E, Switzerland), and an additional 0.5 ml of Trizol was added for a total sample volume of 1.0 ml. 

Real-time PCR
RNA was isolated by the phenol-chloroform method and then quantified using a spectrophotometer (GE healthcare biosciences AB, Uppsala, Sweden). The mean 260/280 absorbance ratio was 1.82���0.05. A 5-�g aliquot of total RNA from each tissue sample was used as a template for reverse transcription and was primed with 1 �l of oligo (dT)18 primer (Clontech, Takara Bio Com., Japan); an appropriate amount of diethyl pyrocarbonate (DEPC)-treated water was added to a final volume of 13.5 �l. The synthesis reaction contained 4 �l of 5X reaction buffer, 1 �l of 10 mM dNTP-mix, 0.5 �l of 40 U/�l recombinant RNase inhibitor, and 1 �l of 200 U/�l MMLV reverse transcriptase in a final volume of 20 �l. The total RNA was preincubated for 2�min at 70�C prior to cDNA synthesis, and the reverse transcription reaction was performed for 60 min at 42�C followed by 5 min at 94�C. The following primer sequences were used to amplify the ChAT, AChE and an internal standard �-actin: �-actin forward primer 52 - AGCCATGTACGTAGCCATCC-32 , reverse primer 52 -TTTGATGTCACGCACGATTT-3, ChAT forward primer 52 -GGTGGCCCAGAAGAGCAGTATC-32 , reverse primer 52 -ATTGGAGGCAGGCGTTCATC-32  and AChE forward primer 52 -TACTCTGGACGAGGCG-32 , reverse primer 52 -CGGTCGTATTATATCCCAGC-32 . A SYBR Green master mix (Roche, USA) was used to measure the accumulation of PCR product for each reaction. The PCR amplification protocol consisted of one cycle at 95 �C for 10 min followed by 45 cycles at the following parameters: 40 s of denaturation at 95 �C, 40 s of annealing at 60 �C and 1 min 20 s of extension at 72 �C using the ABI Prism 7700 Sequence Detection System driven by the ABI prism SDS version 9.1 software (Applied Biosystems). The relative expression of the genes of interest was normalized to the mean of the �-actin (internal control) mRNA that was amplified along with each sample.

Statistics
All values were expressed as the mean�� standard error of the mean (SEM); values of P�<0.05 were considered significant. The student's t-test was used to assess significant differences between the two strains.

3. Results and Discussion
Fig. shows a comparison of ChAT and AChE mRNA expression between ApoE-KO and WT mice. Aldh2-KO mice exhibited significantly lower ChAT expression (P<0.01) than WT mice. No statistically significant strain differences of AChE were observed. Our recent study indicates that ApoE-KO mice have poorer spatial memory [9]. To explore this issue, we compared the hippocampal cholinergic markers expression between ApoE-KO and WT mice. 

The present results demonstrated a significant strain difference in the expression of cholinergic markers between ApoE-KO and WT mice. Specifically, ApoE-KO mice showed decreased ChAT mRNA compared to WT mice, with no difference in AChE mRNA. This later one is in agreement with other work compare AChE between ApoE-KO and WT mice in the regions of the brain showed similar patterns of AChE staining [10,11]. Our finding of low level of ChAT mRNA in ApoE-KO mice was consistent with the previous study that reported that ChAT activity was unchanged in the brain regions studied of ApoE-deficient mice in comparison with WT mice [5]. The synaptic densities of cholinergic and other neural projections in specific brain regions of ApoE-KO mice are markedly lower [12]. Given some previous findings in terms of cholinergic function in ApoE-KO mice [12,13], we have hypothesized that ApoE-KO mice may have a cholinergic deficit. A low level of ChAT can lead to a decrease in ACh, which may play a critical role in cognitive function. Taken together, Apo-E-deficiency related ChAT mRNA decrease in the hippocampus may lead to greater cognitive deficit in Apo-KO mice. 

In conclusion, our data give evidence that mice with ApoE-deficiency have lower expression of ChAT mRNA and suggest a possible association between cholinergic deficit and poorer performance in ApoE-KO mice. 

Acknowledgements
This work was supported in part by the Grant-in-Aid for Scientific Research [Grand No (c) 22590636] from the Ministry of Education, Culture, Sports, Science and Technology, Japan. 
References
1.  HYPERLINK "http://www.scopus.com/authid/detail.url?authorId=7004032365&amp;eid=2-s2.0-0021020898" \o "Show Author Details" Eckenstein F,� HYPERLINK "http://www.scopus.com/authid/detail.url?authorId=7006420825&amp;eid=2-s2.0-0021020898" \o "Show Author Details" Sofroniew MV (1983) Identification of central cholinergic neurons containing both choline acetyltransferase and acetylcholinesterase and of central neurons containing only acetylcholinesterase.  HYPERLINK "http://www.scopus.com/source/sourceInfo.url?sourceId=16764&amp;origin=recordpage" \o "Go to the information page for this source" J Neurosci 3:2286-91.
2.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Mohapel%20P%5BAuthor%5D&cauthor=true&cauthor_uid=15718053" Mohapel P,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Leanza%20G%5BAuthor%5D&cauthor=true&cauthor_uid=15718053" Leanza G,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Kokaia%20M%5BAuthor%5D&cauthor=true&cauthor_uid=15718053" Kokaia M,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Lindvall%20O%5BAuthor%5D&cauthor=true&cauthor_uid=15718053" Lindvall O (2005) Forebrain acetylcholine regulates adult hippocampal neurogenesis and learning.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed/15718053" \o "Neurobiology of aging." Neurobiol Aging 26:939-46.
3. Plassman BL, Welsh-Bohmer KA, Bigler ED, Johnson SC, Anderson CV, Helms MJ, Saunders AM, Breitner JC (1997)  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed/9109888" Apolipoprotein E epsilon 4 allele and hippocampal volume in twins with normal cognition. Neurology 48:985-89.
4.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=%22Greenwood%20PM%22%5BAuthor%5D" Greenwood PM,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=%22Sunderland%20T%22%5BAuthor%5D" Sunderland T,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=%22Friz%20JL%22%5BAuthor%5D" Friz JL,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=%22Parasuraman%20R%22%5BAuthor%5D" Parasuraman R (2000) Genetics and visual attention: selective deficits in healthy adult carriers of the epsilon 4 allele of the apolipoprotein E gene.  HYPERLINK "javascript:AL_get(this,%20'jour',%20'Proc%20Natl%20Acad%20Sci%20U%20S%20A.');" \o "Proceedings of the National Academy of Sciences of the United States of America."  Proc Natl Acad Sci 97:11661-66.
5. Gordon I, Grauer E, Genis I, Sehayek E, Michaelson DM (1995)  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed/8584214?ordinalpos=8&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSum" Memory deficits and cholinergic impairments in apolipoprotein E-deficient mice. Neurosci Lett 199:1-4.
6.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Graybiel%20AM%5BAuthor%5D&cauthor=true&cauthor_uid=1695398" Graybiel AM (1990) Neurotransmitters and neuromodulators in the basal ganglia.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed/1695398?dopt=Abstract" \o "Trends in neurosciences." Trends Neurosci 13:244-54.
7. Holt DJ, Herman MM, Hyde TM, Kleinman JE, Sinton CM, German DC, Hersh LB, Graybiel AM, and Saper CB (1999) Evidence for a deficit in cholinergic interneurons in the striatum in schizophrenia. Neuroscience 94:21-31.
8. Pisani A, Bonsi P, Picconi B, Tolu M, Giacomini P, and Scarnati E (2001) Role of tonically-active neurons in the control of striatal function: cellular mechanisms and behavioral correlates. Prog Neuropsychopharmacol Biol Psychiatry 25:211-30.
9. Jamal M, Ameno K, Miki T, Tanaka N, Ono J, Shirakami G, Sultana R, Yu N, Kinoshita H (2012)  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed/22361447" High ethanol and acetaldehyde impair spatial memory in mouse models: opposite effects of aldehyde dehydrogenase 2 and apolipoprotein E on memory. Pharmacol Biochem Behav 101:443-49. 
10. Anderson R, Higgins GA (1997)  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed/9266610" Absence of central cholinergic deficits in ApoE knockout mice. Psychopharmacology (Berl) 132:135-44.
11.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Krzywkowski%20P%5BAuthor%5D&cauthor=true&cauthor_uid=10426483" Krzywkowski P,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Ghribi%20O%5BAuthor%5D&cauthor=true&cauthor_uid=10426483" Ghribi O,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Gagn%C3%A9%20J%5BAuthor%5D&cauthor=true&cauthor_uid=10426483" Gagn� J,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Chabot%20C%5BAuthor%5D&cauthor=true&cauthor_uid=10426483" Chabot C,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Kar%20S%5BAuthor%5D&cauthor=true&cauthor_uid=10426483" Kar S,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Rochford%20J%5BAuthor%5D&cauthor=true&cauthor_uid=10426483" Rochford J,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Massicotte%20G%5BAuthor%5D&cauthor=true&cauthor_uid=10426483" Massicotte G,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Poirier%20J%5BAuthor%5D&cauthor=true&cauthor_uid=10426483" Poirier J (1999) Cholinergic systems and long-term potentiation in memory-impaired apolipoprotein E-deficient mice.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed/10426483" \o "Neuroscience." Neuroscience 92:1273-86.
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$dha$gd�h$d��d�d@&[$\$a$gd�h$dh�a$gd�h$dh�7$8$H$a$gd�h�C�C�C�CPDQD]D^D`DaD�D�D�D�D�D�DFEGE�E�E�E�E�E�E�E�E�8�9�������������)�*�4�5�7�8�������������������������������堋z�i�z�z hz0h�hCJPJ^J_HEaJ h^lh�hCJPJ^J_HEaJ)jh^lh�hCJPJU^J_HEaJh�hCJPJ^J_HEaJhz0h�h0J1CJaJUh�hCJaJhz0h�h0J5CJaJ$hz0h�h0J>*B*CJaJph�hz0h�hCJaJjhz0h�hCJUaJ) HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed/9453565" Specific neurochemical derangements of brain projecting neurons in apolipoprotein E-deficient mice.J Neurochem 70:708-14.
13.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Kleifeld%20O%5BAuthor%5D&cauthor=true&cauthor_uid=10198822" Kleifeld O,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Diebler%20MF%5BAuthor%5D&cauthor=true&cauthor_uid=10198822" Diebler MF,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Chapman%20S%5BAuthor%5D&cauthor=true&cauthor_uid=10198822" Chapman S,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Oron%20L%5BAuthor%5D&cauthor=true&cauthor_uid=10198822" Oron L,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Michaelson%20DM%5BAuthor%5D&cauthor=true&cauthor_uid=10198822" Michaelson DM (1998) The effects of apolipoprotein E deficiency on brain cholinergic neurons.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed/10198822" \o "International journal of developmental neuroscience : the official journal of the International Society for Developmental Neuroscience." Int J Dev Neurosci 16:755-62.





















Legend
Strain effects on ChAT and AChE mRNA. * P<0.01 versus WT mice. 


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