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David A Freeman

Publications and source records attributed to David A Freeman.

10 recordsLinked to original sources
PubMed2006

The thalamic intergeniculate leaflet mediates locomotor activity-induced reversal of phenotype in photoperiod nonresponsive Siberian hamsters.

The role of the intergeniculate leaflet of the thalamus (IGL) in photoperiod responsiveness was examined in a laboratory-selected line of photoperiod nonresponsive (NR) Siberian hamsters. NR hamsters fail to exhibit typical winter-type responses (i.e., gonadal regression and development of winter-type pelage) when exposed to short day lengths (e.g., 10 h of light/day). Earlier studies revealed that NR hamsters will exhibit winter-type responses when exposed to short photoperiod if they are given free access to a running wheel. The present study tested the hypothesis that this locomotor activity-induced reversal of phenotype is dependent on the IGL. Male NR hamsters underwent destruction of the IGL prior to being housed in short day lengths in cages equipped with running wheels. Activity rhythms were monitored for 8 weeks, after which time pelage response and paired testes weights were obtained. In contrast to sham-operated NR animals given access to running wheels, IGL-ablated animals showed no increase in the duration of nocturnal running wheel activity and became active later in the night than sham-lesioned animals. Lesioned animals also failed to exhibit the typical short photoperiod-induced gonadal regression and pelage molt. The results implicate the IGL in the mechanism by which running wheel activity can influence photoperiodic responses.

Animals↗
PubMed2005

Neuropeptide Y induces torpor-like hypothermia in Siberian hamsters.

Intracerebroventricular (ICV) injections of neuropeptide Y (NPY) are known to decrease body temperature (Tb) of laboratory rats by 1-3 degrees C. Several NPY pathways in the brain terminate in hypothalamic structures involved in energy balance and thermoregulation. Laboratory rats are homeothermic, maintaining Tb within a narrow range. We examined the effect of ICV injected NPY on Tb in the heterothermic Siberian hamster (Phodopus sungorus), a species that naturally undergoes daily torpor in which Tb decreases by as much as 15-20 degrees C. Minimum effective dose was determined in preliminary testing then various doses of NPY were tested in cold-acclimated Siberian hamsters while food was withheld. NPY markedly reduced Tb in the heterothermic Siberian hamster. In addition, the reduction in Tb in 63% of the observations was sufficient to reach the criterion for daily torpor (Tb < 32 degrees C for at least 30 min). Neither the incidence of torpor nor its depth or duration was related to NPY dose. Both likelihood and magnitude of response varied within animals on different test days. NPY decreased 24-h food intake and this was exaggerated in the animals reaching criterion for torpor; the decrease in food intake was positively correlated with the magnitude of the decrease in Tb. The mild hypothermia seen in homeothermic laboratory rats after NPY injected ICV is exaggerated, often greatly, in the heterothermic Siberian hamster. NPY treatment may be activating hypothalamic systems that normally integrate endogenous torpor-producing signals and initiate torpor.

Animals↗
PubMed2004

The thalamic intergeniculate leaflet modulates photoperiod responsiveness in Siberian hamsters.

Siberian hamsters are seasonal breeders that use changes in day length to synchronize their reproductive effort with those times of the year most favorable for successful reproduction. The ability of Siberian hamsters to measure and respond to changes in day length depends upon accurate photoentrainment of the circadian clock in the suprachiasmatic nucleus (SCN) of the hypothalamus. Two pathways have been characterized through which entraining stimuli reach the SCN: the retinohypothalamic tract (RHT), which transmits light information from the retinae, and the geniculohypothalamic tract (GHT) from the intergeniculate leaflet of the thalamus (IGL), which is involved in transmitting both photic and nonphotic cues. Ablating the IGL/GHT results in only modest alterations in entrainment to static day lengths and fails to interfere with seasonal responses induced by transfer from static long day to static short day lengths. Because several studies suggest that the IGL may be involved in tracking the time of dusk and dawn, we sought to determine whether an intact IGL is necessary for hamsters to respond to a simulated natural photoperiod (SNP) in which the time of dusk and dawn gradually changes in a pattern approximating the rate of change in day length that occurs during autumn at the latitude this species inhabits in nature. The results indicate that neurochemical lesions of the IGL alter both the pattern of circadian entrainment and photoperiodic responsiveness of Siberian hamsters to an SNP. Both intact and IGL-lesioned hamsters exhibited testicular regression in shortening day lengths, but only IGL-intact hamsters exhibited seasonal pelage molt.

Animals↗
PubMed2004

Reduced leptin concentrations are permissive for display of torpor in Siberian hamsters.

A photoperiod with a short photophase induces a winterlike phenotype in Siberian hamsters that includes a progressive decrease in food intake and body mass and reproductive organ regression, as well as reversible hypothermia in the form of short-duration torpor. Torpor substantially reduces energy utilization and is not initiated until body mass, fat stores, and serum leptin concentrations are at their nadir. Because photoperiod-dependent torpor is delayed until fat reserves are lowest, leptin concentrations may be a permissive factor for torpor onset. This conjecture was tested by implanting osmotic minipumps into Siberian hamsters manifesting spontaneous torpor; the animals received a constant release of leptin or vehicle for 14 days. Exogenous leptin treatment eliminated torpor in a significant proportion of treated hamsters, whereas treatment with the vehicle did not. Similarly, endogenous serum leptin concentrations were markedly reduced in all animals undergoing daily torpor. Although simply reducing leptin concentrations below a threshold value is not sufficient for torpor initiation, reduced leptin concentrations nevertheless appear necessary for its occurrence. It is proposed that drastically reduced leptin concentrations provide a "starvation signal" to an as yet unidentified central mechanism mediating torpor initiation.

Animals↗
PubMed2004

Validation and implementation of the PowerPlex 16 BIO System STR multiplex for forensic casework.

The PowerPlex 16 BIO multiplex short tandem repeat (STR) system contains the 13 CODIS loci (FGA, TPOX, D8S1179, vWA, D18S51, D21S11, TH01, D3S1358, CSF1PO, D16S539, D7S820, D13S317, and DS5S818), plus two pentanucleotide repeat loci (Penta D and Penta E) and the sex-identifying locus. Amelogenin. The PowerPlex 16 BIO System is optimized for use with the Hitachi FMBIO gel imaging systems. A consortium of seven independent laboratories collaborated to perform the studies defined by the FBI standards for performing a developmental validation, including the evaluation of sample concordance, percent stutter determination, nonprobative casework, precision, sensitivity, mixture determination, effect of substrates, the impact of environmental insults, and species specificity. All samples tested for concordance were consistent except for one sample from the Virginia Division of Forensic Science database that displayed discordance at D13S317, a locus whose primer sequence was altered. Stutter values were comparable to those of other STR multiplex systems, the precision was comparable to other multiplexes analyzed by gel electrophoresis, the DNA profiles were unchanged by the substrate upon which the blood samples were placed, and the nonprobative casework samples re-typed for the PowerPlex 16 BIO System were consistent with previous typing results. When greater than 0.125 ng of DNA was placed into the PowerPlex 16 BIO System amplification reaction, a full profile was generated by all laboratories. The mixture study results were comparable to those reported for other multiplex systems, the environmental study demonstrated a loss of larger molecular weight loci when samples were incubated at elevated temperatures for a prolonged period of time, and the only notable cross species hybridization was observed with primate DNA samples. This extensive validation work performed demonstrates that the PowerPlex 16 BIO System provides STR data of a quality comparable with other PowerPlex STR multiplex kits as well as other widely used STR multiplexes and is thus suitable for evidentiary casework analysis as well as database sample profiling.

Alleles↗
PubMed2002

Absence of pineal-independent mediation of seasonal differences in suprachiasmatic nucleus AVP and VIP mRNA expression in Siberian hamsters.

Assessment of seasonal variations in expression of brain neuropeptide mRNA is complicated by concurrent circadian variations. Because entrainment of suprachiasmatic nucleus (SCN) based rhythms differs in long versus short day lengths, valid seasonal comparisons must be made at equivalent circadian phases. We used a novel experimental design which permitted sampling at identical circadian phases of animals exhibiting opposite seasonal reproductive responses to the same intermediate day length. This allowed us to test whether seasonal changes in arginine vasopressin (AVP) and vasoactive intestinal peptide (VIP) mRNA expression in the SCN occur in the absence of the pineal gland. Juvenile Siberian hamsters were gestated and maintained postnatally in either a long photoperiod (16 h light/day) or short photoperiod (10 h light/day). At the time of weaning (18 days of age), the hamsters were pinealectomized and either transferred to a new photoperiod (10-, 16- or 14-h light/day) or left in the original photoperiod. Hamsters from 10L had substantially smaller and lighter testes than those from 16L. If photoperiodic modulation of AVP and VIP mRNA expression occurs in the absence of the pineal, then transfer of pinealectomized hamsters from a longer (16L) or shorter (10L) photoperiod to an intermediate photoperiod (14L) should result in a differential response with respect to SCN AVP and VIP mRNA expression but not testis size. When sampled at an identical circadian phase (3 h after lights on) in 14L there was no difference in the expression of AVP or VIP mRNA in the SCN between animals previously housed in long versus short day lengths. In contrast to a previous study that did not carefully control for circadian phase, the present findings suggest that seasonal photoperiodic control of SCN neuropeptide mRNA expression depends upon the pineal gland. In addition, the present findings demonstrate a significant, negative correlation between AVP mRNA expression in the SCN and the length of the daily active phase (alpha).

Animals↗
PubMed2002

Periodic arousal from hibernation is necessary for initiation of immune responses in ground squirrels.

Golden-mantled ground squirrels (Spermophilus lateralis) undergo seasonal hibernation during which core body temperature (T(b)) values are maintained 1-2 degrees C above ambient temperature. Hibernation is not continuous. Squirrels arouse at approximately 7-day intervals, during which T(b) increases to 37 degrees C for approximately 16 h; thereafter, they return to hibernation and sustain low T(b)s until the next arousal. Over the course of the hibernation season, arousals consume 60-80% of a squirrel's winter energy budget, but their functional significance is unknown and disputed. Host-defense mechanisms appear to be downregulated during the hibernation season and preclude normal immune responses. These experiments assessed immune function during hibernation and subsequent periodic arousals. The acute-phase response to bacterial lipopolysaccharide (LPS) was arrested during hibernation and fully restored on arousal to normothermia. LPS injection (ip) resulted in a 1-1.5 degrees C fever in normothermic animals that was sustained for > 8 h. LPS was without effect in hibernating squirrels, neither inducing fever nor provoking arousal, but a fever did develop several days later, when squirrels next aroused from hibernation; the duration of this arousal was increased sixfold above baseline values. Intracerebroventricular infusions of prostaglandin E(2) provoked arousal from hibernation and induced fever, suggesting that neural signaling pathways that mediate febrile responses are functional during hibernation. Periodic arousals may activate a dormant immune system, which can then combat pathogens that may have been introduced immediately before or during hibernation.

Animals↗
PubMed2002

Detection of a primer-binding site polymorphism for the STR locus D16S539 using the Powerplex 1.1 system and validation of a degenerate primer to correct for the polymorphism.

Quality assurance samples submitted from the NCSBI as part of a contract with TBTG to outsource DNA Database samples showed unexpected discrepancies for the locus D16S539 when all other loci yielded identical results. Discrepancies observed included allele drop out and an imbalance in sister alleles with samples returned from TBTG. This led to a comprehensive review of the technical procedures used between the two laboratories to determine the cause of the discrepancies noted for the locus D16S539, since both laboratories were using the PowerPlex 1.1 typing kit from the Promega Corporation. The NCSBI and the TBTG utilize different extraction methods (organic extraction vs. FTA) and amplification conditions (AmpliTaq vs AmpliTaq Gold), respectively, so the exact cause of discrepancy observed was not immediately apparent. Experiments at the NCSBI associated the observed allele drop out and the imbalance of the sister alleles with the use of AmpliTaq Gold and a hot start procedure. Sequencing data revealed that a point mutation resides on the D16S539 primer-binding site that reaches polymorphic levels in African-American populations. This led to the development of a degenerate primer by the Promega Corporation to detect "missing" alleles when AmpliTaq Gold is used. The degenerate primer was then thoroughly tested to show its efficacy in detecting the "true" D16S539 profile when used.

Allelic Imbalance↗
PubMed2002

Characterization and validation studies of powerPlex 2.1, a nine-locus short tandem repeat (STR) multiplex system and penta D monoplex.

In order to increase the power of discrimination for human identification purposes, a nine-locus short tandem repeat (STR) multiplex, the GenePrint PowerPlex 2.1 system (PowerPlex 2.1) developed by Promega Corporation and a separate pentanucleotide-repeat locus, Penta D, were tested. This megaplex system includes the highly polymorphic loci FGA, TPOX, D8S1179, vWA, Penta E, D18S51, D21S11, TH01, and D3S1358 and may be used in combination with the eight-locus STR multiplex, the GenePrint PowerPlex 1.1 system (PowerPlex 1.1) that has been previously developed. Three of the loci, TPOX, TH01 and vWA, have been included in both systems for quality control purposes. As with PowerPlex 1.1, PowerPlex 2.1 is also based on a two-color detection of fluorescent-labeled DNA products amplified by polymerase chain reaction (PCR) and provides a valuable tool for accurate and rapid allele determination. The primer sequences used in the PowerPlex 2.1/Penta D system are also presented in this report. To meet the "Quality Assurance Standards for Forensic DNA Testing Laboratories" (FBI), we tested the efficiency and reproducibility of the PowerPlex 2.1/PentaD system by several validation studies that were conducted as a joint project among seven laboratories. Validation tests included concordance studies, sensitivity, and species specificity determination, as well as performance in forensic and environmentally impacted samples. The results produced from these tests demonstrated the consistency and reliability of the PowerPlex 2.1/Penta D system.

Animals↗