Sam DiGiovanni Professor Wolcott ENC 1102 October 21, 2013 Separation Anxiety during the Maternal Phase

Within my major researchers and veterinarians have been talking about separation anxiety occurring to animals at a young age and if it truly affects them as they mature and grow into adults. The way I came about my topic was that within the veterinarian world there are many experiments that occur that use laboratory animals. The most common animals you see being used in the lab are rodents, so every article is an experiment using either mice or rats. The reason why I chose experiments is because the experimenter has the ability to control the procedures and how they will be carried out to gain the best results on the subjects. This annotated bibliography focuses on separation anxiety during the maternal stage where the pups need their mothers the most and what behavior they go through. There are many negative effects, but also there are some positive when separated from their mothers, but the question does it have long-term effects on them? Most of the terms in the articles are medical of scientific. People who could benefit from reading this annotated bibliography are students in pre-vet, teachers, experimenters, veterinarians, or even researchers. Some definitions you may want to know before reading are: Maternal separation (MS)- being apart from the mother Postnatal days (PND)- occurring or being after birth Stressor- something that makes you stressed

Corticosterone- hormone secreted by the adrenal cortex, one of the glucocorticoids.

Alison L. Martin, Richard E. Brown, The lonely mouse: Verification of a separationinduced model of depression in female mice, Behavioural Brain Research, Volume 207, Issue 1, 11 February 2010, Pages 196-207 In this journal article Alison Martin and Richard Brown, researchers from the Psychology Department and Neuroscience Institute, at Dalhousie University, evaluated depression from female mice when they are separated at a young age. They took female mice at 8 weeks of age and separated them from their cage to another until they were 24 weeks old. At 24 weeks of age, there was a significant increase in weight and in immobility in individually housed mice in the forced swim test and tail suspension test, a reduction in transitions in the L/D box, a reduced startle response and reduced pre-pulse inhibition, but no differences in cued or context fear conditioning. (Alison). Another experiment was done where they took female mice at 8 weeks and grouped them together in one cage. From this experiment they found that group-housed females resulted in higher baseline corticosterone (CORT) levels compared to the isolated females in the first experiment and fluoxetine had no effect on CORT levels. Thus, separation from group housing is a reliable and valid method for inducing depression-like behaviour in female mice (Alison). This article shows relevance because in female mice you see they do experience more depression when they are separated from their cage and put into another with other mice of no relation.

Anna Llid, Laura Mdol, Snia Darbra, Marc Pallars, Interaction between neonatal allopregnanolone administration and early maternal separation: Effects on adolescent and adult behaviors in male rat, Hormones and Behavior, Volume 63, Issue 4, April 2013, Pages 577-585 In this journal article Anna Llido and her team of researchers are from Universitat Autonoma de Barcelona, in Spain where they analyzed the interaction between the effects of the neonatal alteration of allopregnanolone (AlloP), neurosteroid that increase its levels after acute stress presentation, and EMS on adolescent exploration and adult anxiety and sensorimotor gating in male rats. (Anna). Results showed that the AlloP neautralized the effects of early maternal separation, which is important because this disrupted the prepulse inhibition that shows neural development and vulnerability to disorders (Anna). Callaghan, Bridget L., and Rick Richardson. "Maternal Separation Results In Early Emergence Of Adult-Like Fear And Extinction Learning In Infant Rats." Behavioral Neuroscience 125.1 (2011): 20-28. Academic Search Premier. Web. 20 Oct. 2013. In this journal article Callaghan and Richardson, researchers from the University of New South Wales, examined the effects of early life stress in developing rats. They started off by separating the rats on postnatal days 2-14 to start an early life stressor and to the rats that didnt get separated were trained to fear a footshock noise (Callaghan). We found that young animals exposed to maternal-separation were more likely to exhibit context and stress-mediated relapse after extinction than the standard-reared rats. (Callaghan). Later on in the experiment the

maternally separated rats had a return of fear when the inhibitory neurotransmitter was blocked when tested, which suggests that early life adversity prepares younger animals to respond to fear in their environment (Callaghan). This article is relevant because early separation for these rats made them grow fear and develop stress, which now is carried with them as they mature. Frdric S. Dumont, Vivian Biancardi, Richard Kinkead, Hypercapnic ventilatory response of anesthetized female rats subjected to neonatal maternal separation: Insight into the origins of panic attacks?, Respiratory Physiology & Neurobiology, Volume 175, Issue 2, 15 February 2011, Pages 288-295 In this journal article Dumont and his team of researchers from Universitaire de Quebec, investigated to understand the mechanisms underlying the sex-specific effects of NMS on the ventilatory response to CO2, we used two different anesthetized female rat preparations to assess central CO2 chemosensitivity and contribution of sensory afferents (stretch receptors and peripheral chemoreceptors) that influence the HCVR. (Frederic). Since the anesthesia did not affect NMS-related respiratory dysfunction in males we propose that the panic or anxiety induced by CO2 during wakefulness is responsible for enhancement of the HCVR in NMS females. (Frederic). This article is important because the anesthesia didnt affect the NMS in males, so the anxiety is coming from the HCVR in the females separation. Henritte J. Hulshof, Arianna Novati, Andrea Sgoifo, Paul G.M. Luiten, Johan A. den Boer, Peter Meerlo, Maternal separation decreases adult hippocampal cell proliferation and impairs cognitive performance but has little effect on stress

sensitivity and anxiety in adult Wistar rats, Behavioural Brain Research, Volume 216, Issue 2, 20 January 2011, Pages 552-560 In this article Hulshof and her team of researchers from the University of Groningen, investigated whether maternal separation or the test/experiments are the ones that are causing the stress on the rats. Rat pups were taken during the postnatal weeks 1-2 and had 3 hours daily of separation. The results show that, independent of the nature of the stressor, MS did not affect the neuroendocrine response, it did not influence anxiety-like behavior, explorative behavior and social interaction, but did affect cognitive function in an object recognition task. (Henriette). This article shows importance because its saying exactly the opposite of what all the other articles are stating, while interfering with the motherinfant relationship early in life did affect some aspects of adult neuroplasticity and cognitive function, it did not lead to permanent changes in stress sensitivity and emotionality. (Henriette). John Cryan, et al. "Maternal Separation As A Model Of Brain-Gut Axis Dysfunction." Psychopharmacology 214.1 (2011): 71-88. Academic Search Premier. Web. 21 Oct. 2013. In this journal article Cryan and his team of researchers from the University College Cork, in Ireland observed that not only does maternal separation cause stress anxiety it also can cause irritable bowel syndrome (IBS) which dysfunctions the interaction between the brain and the gut (Cryan). They claim that evidence suggests that early life stress is a predisposing factor to IBS, so the results show a separated phenotype that is altered by the instinal barrier, altered

balance in microflora, stress response and visceral hypersensitivity which are all cases of IBS (Cryan). This article is important because this shows even more effects on the rats that can happen when they are separated at the maternal stage. Lawrence S. Own, Paresh D. Patel, Maternal behavior and offspring resiliency to maternal separation in c57bl/6 mice, Hormones and Behavior, Volume 63, Issue 3, March 2013, Pages 411-417 In this journal article Own and Patel come from the University of Michigan Medical Center where they investigated the effect of MS on maternal care, offspring behavior, and offspring stress-induced corticosterone response in the c57bl/6 mouse strain. (Lawrence). The results from this study suggest that: (i) levels of maternal care increase as a function of separation duration immediately after daily MS, but long-term care remains unchanged; and (ii) c57bl/6 mice are resilient to MS, exhibiting subtle decreases in anxiety and unchanged stressinduced corticosterone response as adults, irrespective of separation duration. (Lawrence). This article is relevant because it show that the moms show more maternal care when the litter is given back to her showing her anxiety of when they are gone. Len Rodrguez, Diego Armando, and Zulma Dueas. "Maternal Separation During Breastfeeding Induces Gender-Dependent Changes In Anxiety And The GABA-A Receptor Alpha-Subunit In Adult Wistar Rats." Plos ONE 8.6 (2013): 1-8. Academic Search Premier. Web. 21 Oct. 2013. In this journal article Diego and Zulma researchers from Universidad Nacional de Colombia, investigated wistar rats while maternal separation to see whether

breastfeeding affected the males and females anxiety and brain GABA-A receptor-alpha-subunit immunoreactivity (Leon). They housed the rats under a reversed light cycle where they had a light on during certain hours then would turn it off at a certain time. The rats went through separation twice a day during the dark from postnatal day 1-21. Results found were that separated females spent more time in open arms and showed more head dipping than the controls. The separated males spent more time in the center of the maze and engaged in morestrecthing behavior compared to the controls. (Leon). The separation showed gender-specific effects on anxiety behaviors between the females and males, which can be related to the difference GABA-A neurotransmission (Leon). This article is important because the alterations in the GABA-A receptor from not receiving the mothers breastfeeding gives the rats different behaviors of anxiety depending on the gender. L. Zhang, V.S. Hernndez, B. Liu, M.P. Medina, A.T. Nava-Kopp, C. Irles, M. Morales, Hypothalamic vasopressin system regulation by maternal separation: Its impact on anxiety in rats, Neuroscience, Volume 215, 26 July 2012, Pages 135-148 In this journal article Zhang and his team of researchers from the Universidad Nacional Autonoma de Mexico, investigated to see if Arginine vasopressin (AVP) had an effect on anxiety. Male rat pups were separated for three hours daily from postnatal day 2-15 where AVP-mRNA expression was examined within the rats. MS rats demonstrated a high conditioned anxious state under VCT paradigm whereas no difference was found under Elevated plus maze test. (Zhang). This data demonstrated direct relationships between enhanced AVP neuronal activation

and a potentiated vasopressin system, and this latter one with high conditioned anxiety in MS male rats. (Zhang). This is relevant because this showed what arginine vasopressin does to anxiety. Marco Battaglia, et al. "Unstable Maternal Environment, Separation Anxiety, And Heightened CO2 Sensitivity Induced By Gene-By-Environment Interplay." Plos ONE 6.4 (2011): 1-11. Academic Search Premier. Web. 20 Oct. 2013. In this response analysis researchers from Institutes and Universities in Italy hypothesized that early interference with infant-mother interaction could moderate the interspecific trait of response to Carbon dioxide through genetic control of sensitivity to the environment. (1). The researchers used mice for their experiment and had them mate when they were twelve weeks old. Once the pregnant mice had their litters they started their separation methods. They changed caregivers following a rotation scheme presenting the litters to different mothers. This repeated change of caregiver was aimed at interfering with the formation of the infant-mother relationship and to approximate parental instability, a risk factor for internalizing disorders as SAD, PD, Carbon dioxide hypersensitivity. (3). The results on the pups showed that the pups were more vocal, pronounced hyperventilatory responses to Carbon dioxide enriched air, breathing was significantly higher, and received grooming and licking from caregivers. These results show a stress diathesis model where the genetic influences and separation increase Carbon dioxide when a pups environment changes. This response analysis is relevant because within the researchers experiment the mice showed that when they are separated from their mothers

within the maternal stage and are placed in different environments they grow anxiety at an early age. O'Malley, Dervla, Timothy Dinan, and John Cryan. "Neonatal Maternal Separation In The Rat Impacts On The Stress Responsivity Of Central Corticotropin-Releasing Factor Receptors In Adulthood." Psychopharmacology 214.1 (2011): 221-229. Academic Search Premier. Web. 20 Oct. 2013. In this journal article Omalley, Timothy, and Cryan researchers from the University College Cork, Ireland, investigated effects on rats during maternal separation and how it impacts the corticotropin-releasing factor receptors (CRFR). The rats were separated at postnatal day 2-12 and were placed in clean cages with heated blankets until they reached maturity. The results from separation showed that the CRFR1 was elevated in the hypothalamus, pre-frontal cortex and the hippocampus with the addition of the CRFR2 having stress-induced increases in the amygdala of the rats (OMalley). This article is important because when maternal separation occurs it shows how the brain CRF system responds to the stress the rats experience and can lead to persistent changes in central CRF receptor expression and increased sensitivity to stress (OMalley). Tsuda, Mumeko C., and Sonoko Ogawa. "Long-Lasting Consequences Of Neonatal Maternal Separation On Social Behaviors In Ovariectomized Female Mice." Plos ONE 7.3 (2012): 1-11. Academic Search Premier. Web. 21 Oct. 2013. In this journal article Tsuda and Ogawa both researchers from the University of Tsukuba, in Japan examined maternal separation on female social behaviors in different ways. They took the mice from postnatal days 1-14 and put them in

many different environments and showed that MS females preferred to be with other females than male mice. These results suggest that MS females might differ in neuroendocrine responses toward unfamiliar female and male oppents. (Tsuda). This article is important because it shows that early life stress in female mice change their behavior involving social and non-social environments (Tsuda). Vitaly Ryu, Sang Bae Yoo, Dong-Won Kang, Jong-Ho Lee, Jeong Won Jahng, Postweaning isolation promotes food intake and body weight gain in rats that experienced neonatal maternal separation, Brain Research, Volume 1295, 12 October 2009, Pages 127-134 In this journal article Vitaly Ryu and her team of researchers from the Department of Pharmacology, at Kwandong University College of Medicine examined the effects of post-weaning social isolation stress on food intake and body weight gain of rats with maternal separation experience (Vitaly). The rats were separated daily for 180 min during the first 2 weeks of birth. The male pups were caged either in a group of three or singly (social isolation). Maternal separated (MS) pups showed increased immobility in forced swim test and in the elevated plus maze test, group-caged MS pups spent less time in the open arms and more time in the closed arms, but social isolation did not further affect the arm stay of MS pups (Vitaly). These results suggest that post-weaning social isolation may promote hyperphagia and weight gain in young rats that experienced neonatal maternal separation, perhaps, in relation with its impact on the psycho-emotional behaviors of MS pups (Vitaly). This article is important because it shows that

when these young pups were separated and put by themselves they were affected as in they gained weight and had negative behaviors. Wang, Li, Jianwei Jiao, and Stephanie Dulawa. "Infant Maternal Separation Impairs Adult Cognitive Performance In BALB/Cj Mice." Psychopharmacology 216.2 (2011): 207-218. Academic Search Premier. Web. 21 Oct. 2013. In this journal article Wang, Jiao, and Dulawa all researchers at the University of Chicago, examined effects of early adversity on adult emotional behavior, the antidepressant response, and cognitive performance in BALB/cJ mice. (Wang). The split the mice up into separation and controlled groups where the separation mice where from postnatal days 2-15. They received treatment with the serotonin reuptake inhibitor, fluoxetine, and were checked for anxiety or depression. The results showed that IMS reduced fear conditioning to the tone and context, disrupted novel object recognition in females, and impaired both spatial and reversal learning in males. (Wang). This article is relevant because this shows that IMS induces adults emotional, episodic, spatial, and learning memory (Wang). Yoav Litvin, Philip Tovote, Nathan S. Pentkowski, Thomas Zeyda, Lanikea B. King, Amy J. Vasconcellos, Christopher Dunlap, Joachim Spiess, D. Caroline Blanchard, Robert J. Blanchard, Maternal separation modulates short-term behavioral and physiological indices of the stress response, Hormones and Behavior, Volume 58, Issue 2, July 2010, Pages 241-249 In this scholarly journal article Yoav Litvin and other researchers from the University of Hawaii, evaluated rats and their stress responses given off when

separated from their mother during the postnatal stage. The researchers took rat pups during the postnatal stage and put them in a different cage than their mother that had a different odor than their original cage. The results showed maternal separation affected unconditioned freezing on the pups where they didnt move, but other defenses were not triggered, such as avoidance or risk-assessment, showing a tolerant anxiolytic effect (Yoav). There were no sex differences other than weight. We suggest that maternal separation may have produced these changes by disrupting normal development of hippocampal regions involved in olfactory-mediated freezing, not in mechanisms of learning and memory per se. These findings may have direct relevance for understanding the mechanisms by which early-life adverse experiences produce short-term and lasting psychopathologies. This article is relevant because within the experiment the pups froze when being separated from their mothers, which puts stress on them at a young age, which can stay with them as they grow older.