Meng Zhao

All Publications

• Isolation, culture, and functional analysis of hepatocytes from mice with fatty liver disease. STAR protocols Jung, Y., Zhao, M., Svensson, K. J. 2020; 1 (3): 100222

  Abstract

  We present a protocol for isolating hepatocytes from mice with established non-alcoholic fatty liver disease. This protocol consists of liver perfusion using a peristaltic pump, followed by a modified 25% and 90% Percoll gradient centrifugation protocol to capture lipid-laden hepatocytes that are usually lost using traditional isolation protocols. This protocol enables simultaneous isolation of normal and lipid-filled hepatocytes. Lipid-filled hepatocytes can be used in cell culture systems to study drug metabolism, hepatotoxicity, or glucose and lipid metabolism. For complete details on the use and execution of this protocol, please refer to Sharabi etal. (2017) and Kegel etal. (2016).

  View details for DOI 10.1016/j.xpro.2020.100222

  View details for PubMedID 33377114

• Effects of sex and age on parental motivation in adult virgin California mice BEHAVIOURAL PROCESSES Nguyen, C. Y., Zhao, M., Saltzman, W. 2020; 178: 104185

  Abstract

  Female mammals often demonstrate a rapid initiation of maternal responsiveness immediately after giving birth, as a result of neuroendocrine changes that occur during pregnancy and parturition. However, fathers and virgins of some species may display infant care similar to that performed by mothers but without experiencing these physiological events. In biparental species, in which both mothers and fathers care for their offspring, both sex and age may affect parental motivation, even in adult virgins. We examined the effects of sex and age on parental motivation in the California mouse, a monogamous, biparental rodent. We compared parental motivation of male and female virgins in both mid- and old adulthood using two new tests - a T-maze test and a rain test - as well as in standard parental-behavior tests. Adult virgin males were more parentally motivated than adult virgin females in both the T-maze test and the parental-behavior test, but parental motivation did not differ markedly between middle-aged and older adults of either sex. These findings suggest that sex differences in parental motivation in adult virgins are similar to those observed in other biparental rodents, and indicate that the T-maze test may be useful for evaluating parental motivation in this species.

  View details for DOI 10.1016/j.beproc.2020.104185

  View details for Web of Science ID 000555776300013

  View details for PubMedID 32603677

• Long-Term Effects of Fatherhood on Morphology, Energetics, and Exercise Performance in California Mice (Peromyscus californicus) PHYSIOLOGICAL AND BIOCHEMICAL ZOOLOGY Andrew, J. R., Garland, T., Chappell, M. A., Zhao, M., Horrell, N. D., Saltzman, W. 2020; 93 (1): 75–86

  Abstract

  In male mammals that provide care for their offspring, fatherhood can lead to changes in behavioral, morphological, and physiological traits, some of which might constitute trade-offs. However, relatively little is known about these changes, especially across multiple reproductive bouts, which are expected to magnify differences between fathers and nonreproductive males. We evaluated consequences of fatherhood in the monogamous, biparental California mouse (Peromsycus californicus) across seven consecutive reproductive bouts. We compared breeding adult males (housed with sham-ovariectomized females) with two control groups: nonbreeding males (housed with ovariectomized females treated with estrogen and progesterone to induce estrous behavior) and virgin males (housed with untreated ovariectomized females). At five time points (before pairing, early postpartum of the first litter, late postpartum of the second litter, early postpartum of the sixth litter, and late postpartum of the seventh litter or comparable time points for nonbreeding and virgin males), we measured males' body composition, hematocrit, predatory aggression, resting metabolic rate, maximal oxygen consumption ( V ˙ O 2  max ⁡ ), grip strength, and sprint speed. We also weighed organs at the final time point. We predicted that fathers would have lower relative body fat and lower performance abilities compared with control groups and that these effects would become more pronounced with increasing parity. Contrary to predictions, breeding and control males differed in surprisingly few measures, and the number and magnitude of differences did not increase with parity. Thus, our expectations regarding trade-offs were not met. As reported in studies of single reproductive events, these results suggest that fatherhood has few costs in this species when housed under standard laboratory conditions, even across multiple reproductive bouts.

  View details for DOI 10.1086/706863

  View details for Web of Science ID 000501783200002

  View details for PubMedID 31808736

• Regulation of Energy Metabolism by Receptor Tyrosine Kinase Ligands. Frontiers in physiology Zhao, M., Jung, Y., Jiang, Z., Svensson, K. J. 2020; 11: 354

  Abstract

  Metabolic diseases, such as diabetes, obesity, and fatty liver disease, have now reached epidemic proportions. Receptor tyrosine kinases (RTKs) are a family of cell surface receptors responding to growth factors, hormones, and cytokines to mediate a diverse set of fundamental cellular and metabolic signaling pathways. These ligands signal by endocrine, paracrine, or autocrine means in peripheral organs and in the central nervous system to control cellular and tissue-specific metabolic processes. Interestingly, the expression of many RTKs and their ligands are controlled by changes in metabolic demand, for example, during starvation, feeding, or obesity. In addition, studies of RTKs and their ligands in regulating energy homeostasis have revealed unexpected diversity in the mechanisms of action and their specific metabolic functions. Our current understanding of the molecular, biochemical and genetic control of energy homeostasis by the endocrine RTK ligands insulin, FGF21 and FGF19 are now relatively well understood. In addition to these classical endocrine signals, non-endocrine ligands can govern local energy regulation, and the intriguing crosstalk between the RTK family and the TGFβ receptor family demonstrates a signaling network that diversifies metabolic process between tissues. Thus, there is a need to increase our molecular and mechanistic understanding of signal diversification of RTK actions in metabolic disease. Here we review the known and emerging molecular mechanisms of RTK signaling that regulate systemic glucose and lipid metabolism, as well as highlighting unexpected roles of non-classical RTK ligands that crosstalk with other receptor pathways.

  View details for DOI 10.3389/fphys.2020.00354

  View details for PubMedID 32372975

  View details for PubMedCentralID PMC7186430

• Effects of short- and long-term cold acclimation on morphology, physiology, and exercise performance of California mice (Peromyscus californicus): potential modulation by fatherhood JOURNAL OF COMPARATIVE PHYSIOLOGY B-BIOCHEMICAL SYSTEMS AND ENVIRONMENTAL PHYSIOLOGY Andrew, J. R., Garland, T., Chappell, M. A., Zhao, M., Saltzman, W. 2019; 189 (3-4): 471–87

  View details for DOI 10.1007/s00360-019-01219-7

  View details for Web of Science ID 000477891200011

• Effects of single parenthood on mothers' behavior, morphology, and endocrine function in the biparental California mouse. Hormones and behavior Zhao, M., Harris, B. N., Nguyen, C. T., Saltzman, W. 2019; 114: 104536

  Abstract

  Motherhood is energetically costly for mammals and is associated with pronounced changes in mothers' physiology, morphology and behavior. In ~5% of mammals, fathers assist their mates with rearing offspring and can enhance offspring survival and development. Although these beneficial consequences of paternal care can be mediated by direct effects on offspring, they might also be mediated indirectly, through beneficial effects on mothers. We tested the hypothesis that fathers in the monogamous, biparental California mouse (Peromyscus californicus) reduce the burden of parental care on their mates, and therefore, that females rearing offspring with and without assistance from their mates will show differences in endocrinology, morphology and behavior, as well as in the survival and development of their pups. We found that pups' survival and development in the lab did not differ between those raised by a single mother and those reared by both mother and father. Single mothers spent more time in feeding behaviors than paired mothers. Both single and paired mothers had higher lean mass and/or lower fat mass and showed more anxiety-like behavior in open-field tests and tail-suspension tests, compared to non-breeding females. Single mothers had higher body-mass-corrected liver and heart masses, but lower ovarian and uterine masses, than paired mothers and/or non-breeding females. Mass of the gastrointestinal tract did not differ between single and paired mothers, but single mothers had heavier gastrointestinal tract compared to non-breeding females. Single motherhood also induced a flattened diel corticosterone rhythm and a blunted corticosterone response to stress, compared to non-breeding conditions. These findings suggest that the absence of a mate induces morphological and endocrine changes in mothers, which might result from increased energetic demands of pup care and could potentially help maintain normal survival and development of pups.

  View details for DOI 10.1016/j.yhbeh.2019.05.005

  View details for PubMedID 31153926

• Behavioral and endocrine consequences of placentophagia in male California mice (Peromyscus californicus) PHYSIOLOGY & BEHAVIOR Perea-Rodriguez, J. P., Zhao, M., Harris, B. N., Raqueno, J., Saltzman, W. 2018; 188: 283–90

  Abstract

  Ingestion of placenta by mammalian mothers can lead to changes in pain sensitivity, hormone levels, and behavioral responses to newborns. In some biparental mammals, males, in addition to females, ingest placenta when their offspring are born. In the monogamous, biparental California mouse (Peromyscus californicus), males first become attracted to placenta when cohabitating with their pregnant mate, and virgin males administered placenta are less neophobic than males given oil vehicle. In this study, we investigated the effects of placentophagia on pain sensitivity, anxiety-like behavior, behavioral responses to pups, and circulating corticosterone levels of both breeding and nonbreeding male California mice. We orally administered either a conspecific placenta or oil vehicle to male mice from three reproductive conditions (first-time fathers, first-time expectant fathers, and virgin males) and tested their pain sensitivity 1 h later, as well as their exploratory behavior and paternal responsiveness in an open field 4 h post-treatment. We measured plasma corticosterone immediately after the open-field test. We found that placenta-treated males, independent of reproductive condition, traveled significantly longer distances in the open field than males treated with oil, indicative of lower anxiety. Additionally, fathers had shorter latencies to approach and to care for pups (i.e., huddling and licking pups), and spent more time engaging in these behaviors, than did age-matched expectant fathers and virgin males, independent of treatment. We found no effect on plasma corticosterone levels or pain sensitivity as a result of either treatment or reproductive condition. These findings indicate that placenta ingestion decreases anxiety-related behaviors in male California mice, but might not influence pain sensitivity, paternal responsiveness, or plasma corticosterone concentrations.

  View details for PubMedID 29452152

• Effects of a physical and energetic challenge on male California mice (Peromyscus californicus): modulation by reproductive condition JOURNAL OF EXPERIMENTAL BIOLOGY Zhao, M., Garland, T., Chappell, M. A., Andrew, J. R., Harris, B. N., Saltzman, W. 2018; 221 (1)

  Abstract

  Reproduction strongly influences metabolism, morphology and behavior in female mammals. In species in which males provide parental care, reproduction might have similar effects on fathers. We examined effects of an environmental challenge on metabolically important physiological, morphological and behavioral measures, and determined whether these effects differed between reproductive and non-reproductive males in the biparental California mouse (Peromyscus californicus). Males were paired with an ovary-intact female, an ovariectomized female treated with estrogen and progesterone to induce estrus, or an untreated ovariectomized female. Within each group, half of the animals were housed under standard laboratory conditions and half in cages requiring them to climb wire towers to obtain food and water; these latter animals were also fasted for 24 h every third day. We predicted that few differences would be observed between fathers and non-reproductive males under standard conditions, but that fathers would be in poorer condition than non-reproductive males under challenging conditions. Body and fat mass showed a housing condition×reproductive group interaction: the challenge condition increased body and fat mass in both groups of non-reproductive males, but breeding males were unaffected. Males housed under the physical and energetic challenge had higher blood lipid content, lower maximal aerobic capacity and related traits (hematocrit and relative triceps surae mass), increased pain sensitivity and increased number of fecal boli excreted during tail-suspension tests (a measure of anxiety), compared with controls. Thus, our physical and energetic challenge paradigm altered metabolism, morphology and behavior, but these effects were largely unaffected by reproductive condition.

  View details for PubMedID 29170256

• Paternal Care in Biparental Rodents: Intra- and Inter-individual Variation Saltzman, W., Harris, B. N., De Jong, T. R., Perea-Rodriguez, J. P., Horrell, N. D., Zhao, M., Andrew, J. R. OXFORD UNIV PRESS INC. 2017: 589–602

  Abstract

  Parental care by fathers, although rare among mmmals, can be essential for the survival and normal development of offspring in biparental species. A growing body of research on biparental rodents has identified several developmental and experiential influences on paternal responsiveness. Some of these factors, such as pubertal maturation, interactions with pups, and cues from a pregnant mate, contribute to pronounced changes in paternal responsiveness across the course of the lifetime in individual males. Others, particularly intrauterine position during gestation and parental care received during postnatal development, can have long-term effects on paternal behavior and contribute to stable differences among individuals within a species. Focusing on five well-studied, biparental rodent species, we review the developmental and experiential factors that have been shown to influence paternal responsiveness, and consider their roles in generating both intra- and inter-individual variation. We also review hormones and neuropeptides that have been shown to modulate paternal care and discuss their potential contributions to behavioral differences within and between males. Finally, we discuss the possibility that vasopressinergic and possibly oxytocinergic signaling within the brain, modulated by gonadal steroid hormones, may represent the "final common pathway" mediating effects of developmental and experiential variables on intra- and inter-individual variation in paternal care.

  View details for PubMedID 28641377

• Metabolic and and affective consequences of fatherhood in male California mice PHYSIOLOGY & BEHAVIOR Zhao, M., Garland, T., Chappell, M. A., Andrew, J. R., Saltzman, W. 2017; 177: 57–67

  Abstract

  Physiological and affective condition can be modulated by the social environment and parental state in mammals. However, in species in which males assist with rearing offspring, the metabolic and affective effects of pair bonding and fatherhood on males have rarely been explored. In this study we tested the hypothesis that fathers, like mothers, experience energetic costs as well as behavioral and affective changes (e.g., depression, anxiety) associated with parenthood. We tested this hypothesis in the monogamous, biparental California mouse (Peromyscus californicus). Food intake, blood glucose and lipid levels, blood insulin and leptin levels, body composition, pain sensitivity, and depression-like behavior were compared in males from three reproductive groups: virgin males (VM, housed with another male), non-breeding males (NB, housed with a tubally ligated female), and breeding males (BM, housed with a female and their first litter). We found statistically significant (P<0.007, when modified for Adaptive False Discovery Rate) or nominally significant (0.007

  View details for DOI 10.1016/j.physbeh.2017.04.010

  View details for Web of Science ID 000403993900009

  View details for PubMedID 28414073

  View details for PubMedCentralID PMC5500218

• Hormones and the Evolution of Complex Traits: Insights from Artificial Selection on Behavior INTEGRATIVE AND COMPARATIVE BIOLOGY Garland, T., Zhao, M., Saltzman, W. 2016; 56 (2): 207–24

  Abstract

  Although behavior may often be a fairly direct target of natural or sexual selection, it cannot evolve without changes in subordinate traits that cause or permit its expression. In principle, changes in endocrine function could be a common mechanism underlying behavioral evolution because they are well positioned to mediate integrated responses to behavioral selection. More specifically, hormones can influence both motivational (e.g., brain) and performance (e.g., muscles) components of behavior simultaneously and in a coordinated fashion. If the endocrine system is often "used" as a general mechanism to effect responses to selection, then correlated responses in other aspects of behavior, life history, and organismal performance (e.g., locomotor abilities) should commonly occur because any cell with appropriate receptors could be affected. Ways in which behavior coadapts with other aspects of the phenotype can be studied directly through artificial selection and experimental evolution. Several studies have targeted rodent behavior for selective breeding and reported changes in other aspects of behavior, life history, and lower-level effectors of these organismal traits, including endocrine function. One example involves selection for high levels of voluntary wheel running, one aspect of physical activity, in four replicate High Runner (HR) lines of mice. Circulating levels of several hormones (including insulin, testosterone, thyroxine, triiodothyronine) have been characterized, three of which-corticosterone, leptin, and adiponectin-differ between HR and control lines, depending on sex, age, and generation. Potential changes in circulating levels of other behaviorally and metabolically relevant hormones, as well as in other components of the endocrine system (e.g., receptors), have yet to be examined. Overall, results to date identify promising avenues for further studies on the endocrine basis of activity levels.

  View details for DOI 10.1093/icb/icw040

  View details for Web of Science ID 000381279100009

  View details for PubMedID 27252193

  View details for PubMedCentralID PMC5964798